Cell Signaling Technology品牌代理商

上海金畔生物科技有限公司是Cell Signaling Technology品牌代理商 ,欢迎访问官网了解更多产品信息和订购。
Cell Signaling Technology

简要描述:

Cell Signaling Technology, Inc (CST) 是美国著名的生物公司和细胞信号转导研究的,提供特色的信号转导检测用抗体及磷酸化抗体、ELISA试剂盒、激酶等产品。

Cell Signaling Technology, Inc (CST) 是美国著名的生物公司和细胞信号转导研究的,提供特色的信号转导检测用抗体及磷酸化抗体、ELISA试剂盒、激酶等产品。CST总部位于美国波士顿,毗邻哈佛、麻省理工等著名学府的雄厚人才资源,拥有专业的生产和研发队伍,发表的文章多次收录在Nature, Cell, Journal of Immunology 等杂志上。其高质量的产品和专业的研发精神已被全球客户推崇为“细胞信号转导研究的金标准”!

Cell Signaling Technology (CST) is a private, family-owned company, founded by scientists and dedicated to providing the world’s highest quality, innovative research and diagnostic products to accelerate biological understanding and enable personalized medicine. Our employees operate worldwide from our U.S. headquarters in Massachusetts, and our offices in the Netherlands, China, and Japan.

产品列表:

No.

品牌

货号

名称

规格

1

cst

9601

Phospho-(Ser) 14-3-3 Binding Motif Antibody

100 µl

2

cst

9606S

Phospho-(Ser) 14-3-3 Binding Motif (4E2) Mouse mAb

100 µl

3

cst

5522S

14-3-3γ(D15B7) Rabbit mAb

100 µl

4

cst

9769T

14-3-3 Family Antibody Sampler Kit 

1kit

5

cst

9640S

14-3-3ηAntibody

100ul

6

cst

2855T

Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb

20ul

7

cst

6592S

Annexin V-FITC Early Apoptosis Detection Kit

1kit

8

cst

5492S

BrdU Cell Proliferation Chemiluminescent Assay Kit

1kit

9

cst

6813S

BrdU Cell Proliferation Assay Kit

1kit

10

cst

5723S

Caspase-3 Activity Assay Kit

1kit

11

cst

8020S

Cyclic GMP XP® Chemiluminescent Assay Kit

1kit

12

cst

5939S

AIF (D39D2) XP® Rabbit mAb (Sepharose® Bead Conjugate)

400ul

13

cst

8050S

Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb (Magnetic Bead Conjugate)

400ul

14

cst

9646S

Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb (Sepharose® Bead Conjugate)

400ul

15

cst

9563S

Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Magnetic Bead Conjugate) 

400ul

Cell Biolabs特约代理

简要描述:

Cell Biolabs特约代理货品多且全,目前,以我们美国公司为主体,与欧美100多个品牌建立了正式的代理与合作,基本涵盖所有的生物试剂门类,特别是二三线高质量品牌产品,优势更大!

Cell Biolabs是美国著名细胞学试剂品牌,作为前沿的细胞学分析检测试剂盒生产商,为生物技术和研究类客户提供基于细胞功能研究以及疾病等方面的产品和技术服务。Cell Biolabs的诸多产品均采用了技术,比如病毒纯化方面,其开发的病毒吸附Matrix能有效的识别不同的病毒包膜蛋白而特异吸附不同的病毒颗粒。另外,其Cell Biolabs拥有的Platinum (Plat-E, Plat-A and Plat-GP) 逆转录病毒包装细胞系,在生长稳定性和病毒包装滴度方面表现优异,成为众多逆转录病毒包装细胞系中的细胞。

Cell Biolabs, Inc. proudly develops and commercializes innovative technologies and tools for life science research. We are committed to providing the best products that facilitate discovery of the mechanisms underlying cell function and disease.

Our unique, cutting-edge products are currently used around the world in the research laboratories of universities, government institutes, biotechnology and pharmaceutical companies.

Cell Biolabs特约代理产品列表:

cell biolabs

AAV-200

ViraDuctin™ AAV Transduction Kit (10 Transductions)

10 transductions

cell biolabs

AAV-201

ViraDuctin™ AAV Transduction Kit (50 Transductions)

50 transductions

cell biolabs

AAV-300

AAV2 Null Control Virus

50 μL

cell biolabs

AAV-301

AAV1-GFP Control Virus

50 μL

cell biolabs

AAV-302

AAV2-GFP Control Virus

50 μL

cell biolabs

AAV-303

AAV3-GFP Control Virus

50 μL

cell biolabs

AAV-305

AAV5-GFP Control Virus

50 μL

cell biolabs

AAV-306

AAV6-GFP Control Virus

50 μL

cell biolabs

AAV-310

AAV2-Cre Control Virus

50 μL

cell biolabs

AAV-320

AAV2-Luc Control Virus

50 μL

cell biolabs

AAV-400

pAAV-GFP Control Vector

10  µg

cell biolabs

AAV-401

pAAV-Cre Control Vector

10  µg

cell biolabs

AAV-402

pAAV-LacZ Control Vector

10  µg

cell biolabs

AAV-410

pscAAV-GFP Control Vector

10  µg

cell biolabs

AD-200

ViraDuctin™ Adenovirus Transduction Kit (10 Transductions)

10 transductions

cell biolabs

AD-201

ViraDuctin™ Adenovirus Transduction Kit (50 Transductions)

50 transductions

cell biolabs

ADV-001

Null Control Recombinant Adenovirus

50 μL

cell biolabs

ADV-002

β-Galalactosidase Recombinant Adenovirus

50 μL

cell biolabs

ADV-003

SEAP (Secretory Alkaline Phosphatase) Recombinant Adenovirus

50 μL

cell biolabs

ADV-004

GFP Recombinant Adenovirus

50 μL

cell biolabs

ADV-005

Cre Recombinant Adenovirus

50 μL

cell biolabs

ADV-008

Firefly Luciferase Recombinant Adenovirus

50 μL

cell biolabs

ADV-100

Hypoxia Inducible Factor-1 alpha Recombinant Adenovirus

50 μL

cell biolabs

ADV-101

VEGF Recombinant Adenovirus

50 μL

cell biolabs

ADV-103

Interferon γ Recombinant Adenovirus

50 μL

cell biolabs

ADV-104

p38α Recombinant Adenovirus

50 μL

cell biolabs

ADV-105

p38α (Dominant Negative) Recombinant Adenovirus

50 μL

cell biolabs

ADV-112

ERK2 Recombinant Adenovirus

50 μL

cell biolabs

ADV-113

ERK2 (Dominant Negative) Recombinant Adenovirus

50 μL

cell biolabs

ADV-114

JNK1 Recombinant Adenovirus

50 μL

cell biolabs

ADV-115

JNK1 (Dominant Negative) Recombinant Adenovirus

50 μL

cell biolabs

ADV-118

MEK1 (Dominant Negative) Recombinant Adenovirus

50 μL

cell biolabs

ADV-119

MEK1 (Constitutively Active) Recombinant Adenovirus

50 μL

cell biolabs

ADV-120

MKK3 Recombinant Adenovirus

50 μL

cell biolabs

ADV-121

MKK3 (Dominant Negative) Recombinant Adenovirus

50 μL

cell biolabs

ADV-122

MKK3 (Constitutively Active) Recombinant Adenovirus

50 μL

cell biolabs

ADV-123

MKK6 Recombinant Adenovirus

50 μL

cell biolabs

ADV-124

MKK6 (Dominant Negative) Recombinant Adenovirus

50 μL

cell biolabs

ADV-125

MKK6 (Constitutively Active) Recombinant Adenovirus

50 μL

cell biolabs

ADV-126

MKK7 Recombinant Adenovirus

50 μL

cell biolabs

ADV-127

MKK7 (Dominant Negative) Recombinant Adenovirus

50 μL

cell biolabs

ADV-128

MKK7 (Constitutively Active) Recombinant Adenovirus

50 μL

cell biolabs

ADV-129

MEK5 Recombinant Adenovirus

50 μL

cell biolabs

ADV-130

MEK5 (Dominant Negative) Recombinant Adenovirus

50 μL

cell biolabs

ADV-131

MEK5 (Constitutively Active) Recombinant Adenovirus

50 μL

cell biolabs

ADV-132

Raf 1 Recombinant Adenovirus

50 μL

cell biolabs

ADV-133

Raf1 (Dominant Negative) Recombinant Adenovirus

50 μL

cell biolabs

ADV-134

Raf1 (constitutively active) Recombinant Adenovirus

50 μL

cell biolabs

ADV-135

MEKK1 Recombinant Adenovirus

50 μL

cell biolabs

ADV-136

MEKK1 (Dominant Negative) Recombinant Adenovirus

50 μL

cell biolabs

ADV-137

MAPKAPK2 Recombinant Adenovirus

50 μL

cell biolabs

ADV-138

MAPKAPK2 (Dominant Negative) Recombinant Adenovirus

50 μL

cell biolabs

ADV-139

MAPKAPK2 (Constitutively Active) Recombinant Adenovirus

50 μL

cell biolabs

ADV-145

Ras N17 (Dominant Negative) Recombinant Adenovirus

50 μL

 

Premix WST-1 Cell Proliferation Assay System酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Premix WST-1 Cell Proliferation Assay System
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Takara MK400 Premix WST-1 Cell Proliferation Assay System 2,500 次 ¥3,405 Premix WST-1 Cell Proliferation Assay System Premix WST-1 Cell Proliferation Assay System Premix WST-1 Cell Proliferation Assay System
收藏产品 加入购物车
 
■ 制品内容
Premix WST-1 25 ml
 
■ 制品说明
本制品是通过吸光度比色分析法对细胞增殖和细胞活力进行检测的试剂。其检测原理是利用活细胞线粒体中的琥珀酸四唑盐还原酶 (succinate-tetrazolium reductase) 将四唑盐WST-1还原为水溶性formazan dye,进而通过吸光度比色分析法进行检测。此方法取代了[3H]-thymide掺入法,实现了非放射性检测。琥珀酸四唑盐还原酶存在于线粒体中,只有在活细胞中才具有活性,培养基中添加的四唑盐在琥珀酸四唑盐还原酶的作用下,还原生成formazan dye,活细胞数越多,样品中的琥珀酸四唑盐还原酶活性就越强,生成的formazan dye就越多,生成的formazan dye数量与活细胞数成线性正比关系。使用本制品检测时,无需洗涤细胞和使用放射性同位素,从微量培养开始,即可使用酶标仪在同一个微孔板上测定其光吸收值。本制品通过检测细胞活力,可以用于药物敏感性分析和药物筛选。
 
■ 保存
-20℃避光保存。
 
■ 特点
◇ 无需使用放射性同位素。
◇ 无需使用有机溶剂来溶解反应生成的formazan dye。
◇ 吸光度值与活细胞数成线性正比关系。
◇ 与MTT法相比,灵敏度更高。
◇ 可在同一个微孔板上进行不同反应时间的结果测定。
 
■ 用途
◇ 检测生长因子、cytokines、mitogens、nutrients等细胞增殖能力的影响。
◇ 分析抗癌药等具有细胞毒性和抑制细胞生长的化合物。
◇ 评估具有抑制细胞生长作用的抗体与生理介质。
 
 

Premix WST-1 Cell Proliferation Assay System酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Premix WST-1 Cell Proliferation Assay System
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Takara MK400 Premix WST-1 Cell Proliferation Assay System 2500 Tests ¥3,405 Premix WST-1 Cell Proliferation Assay System Premix WST-1 Cell Proliferation Assay System Premix WST-1 Cell Proliferation Assay System
收藏产品 加入购物车
 
■ 制品内容
Premix WST-1 25 ml
 
■ 制品说明
本制品是通过吸光度比色分析法对细胞增殖和细胞活力进行检测的试剂。其检测原理是利用活细胞线粒体中的琥珀酸四唑盐还原酶 (succinate-tetrazolium reductase) 将四唑盐WST-1还原为水溶性formazan dye,进而通过吸光度比色分析法进行检测。此方法取代了[3H]-thymide掺入法,实现了非放射性检测。琥珀酸四唑盐还原酶存在于线粒体中,只有在活细胞中才具有活性,培养基中添加的四唑盐在琥珀酸四唑盐还原酶的作用下,还原生成formazan dye,活细胞数越多,样品中的琥珀酸四唑盐还原酶活性就越强,生成的formazan dye就越多,生成的formazan dye数量与活细胞数成线性正比关系。使用本制品检测时,无需洗涤细胞和使用放射性同位素,从微量培养开始,即可使用酶标仪在同一个微孔板上测定其光吸收值。本制品通过检测细胞活力,可以用于药物敏感性分析和药物筛选。
 
■ 保存
-20℃避光保存。
 
■ 特点
◇ 无需使用放射性同位素。
◇ 无需使用有机溶剂来溶解反应生成的formazan dye。
◇ 吸光度值与活细胞数成线性正比关系。
◇ 与MTT法相比,灵敏度更高。
◇ 可在同一个微孔板上进行不同反应时间的结果测定。
 
■ 用途
◇ 检测生长因子、cytokines、mitogens、nutrients等细胞增殖能力的影响。
◇ 分析抗癌药等具有细胞毒性和抑制细胞生长的化合物。
◇ 评估具有抑制细胞生长作用的抗体与生理介质。
 
 

Luna® Cell Ready 一步法 RT-qPCR 试剂盒 货 号 #E3030S-NEB酶试剂 New England Biolabs

上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。

产品资料 – DNA聚合酶与扩增技术 – qPCR & RT-qPCR

Luna® Cell Ready 一步法 RT-qPCR 试剂盒                              收藏

货 号
规 格
价 格(元)
北京库存
上海库存
广州库存
成都库存
苏州库存

#E3030S
100 rxns
11,669.00

Download:       

  • isoschizomers     |
  • compatible ends     | 
  • single letter code

特性

无需纯化,直接从细胞到 RNA 定量

•  试剂盒包含 100 次裂解反应和 500 次染料一步法 RT-qPCR 反应

   有效裂解多种细胞系的 10 10 万个细胞

   仅需 15 分钟即可完成细胞裂解、RNA 释放和基因组 DNA 去除

  联合使用 LunaWarmStart® RT Hot Start Taq,提高了热稳定性,且可在室温下建立反应体系

   可单独购买裂解模块:Luna Cell Ready 裂解模块(NEBE3032

概述

 Luna Cell Ready 染料一步法 RT-qPCR 试剂盒为直接染料法 RNA 检测和定量提供了所有必要的组分,且无需进行 RNA 提取和纯化。Luna Cell Ready 染料一步法 RT-qPCR 试剂盒包括:1Luna Cell Ready 裂解模块(NEBE3032S)和 2Luna 通用一步法 RT-qPCR KitNEBE3005L)。

细胞培养常用于替代活体生物来分析基因表达或对治疗的反应。传统上使用柱提法或化学试剂法从处理过的细胞中提取和纯化 RNA

 

Luna Cell Ready 裂解模块整合了 DNase I Luna Cell Ready 蛋白酶的功能,仅需 15 分钟即可完成细胞裂解、RNA 释放和基因组 DNA 去除。裂解模块包含独特的 Luna Cell Ready RNA 保护试剂,可在细胞裂解过程中保持 RNA 完整性。50 µl 裂解体系下可对 10-10 万个细胞进行裂解反应。2 µl 裂解产物 (相当于 0.2–4000 个细胞的 RNA 即可实现 20 µl 体系的 RT-qPCR 反应。与其它 Luna 产品一样,裂解缓冲液包含惰性蓝色示踪染料,使整个加样流程可视化。 

Luna 通用一步法 RT-qPCR 试剂盒可兼容大多数实时荧光定量仪器的 SYBR/FAM 荧光通道,进行目标 RNA 的实时定量分析。在该试剂盒中,联合使用热启动 Taq DNA 聚合酶与新型 Luna WarmStart 反转录酶,通过适配体可逆抑制来双重控制酶活性。Luna 通用一步法 RT-qPCR 试剂盒可以直接将 Luna Cell Ready 裂解模块制备的细胞裂解物中的 RNA 转录本进行检测和定量。 

下表列出的细胞系可使用 Luna Cell Ready 裂解模块裂解。这些细胞进行 5 log 梯度稀释(100000-10 细胞/50 µl 裂解反应),并取 1 ul 裂解产物进行 20 µl 体系的一步法 RT-qPCR 反应。每个细胞系的线性结果区域显示在最后一列。此外,经测试一些昆虫细胞系也能用上述裂解液裂解。

 

表一:经验证过的细胞系

 

细胞系

 

特性

菌种

50 ml 裂解体系中的细胞数量

A549

Adherent

H. sapiens, Lung, carcinoma

10 – 100,000

HEK293

Adherent

H. sapiens, Kidney

10 – 100,000

HeLa

Adherent

H. sapiens, Cervix, adenocarcinoma

10 – 100,000

HepG2

Adherent

H. sapiens, Liver, carcinoma

10 – 100,000

NCI-H460

Adherent

H. sapiens, Lung, carcinoma

10 – 100,000

SK-N-SH

Adherent

H. sapiens, Brain, Neuroblastoma

10 – 100,000

U2Os

Adherent

H. sapiens, Bone, osteosarcoma

10 – 10,000

Jurkat

Suspension

H. sapiens, T lymphocyte, leukemia

10 – 100,000

K-562

Suspension

H. sapiens, Lymphoblast, leukemia

10 – 1,000

 

图一:Luna Cell Ready 一步法 RT-qPCR 实验流程

Luna® Cell Ready 一步法 RT-qPCR 试剂盒            货   号                  #E3030S

Luna Cell Ready 一步法 RT-qPCR 试剂盒提供了直接从细胞(50 µl 体系可裂解高达 10 个细胞)到 RNA 检测和定量的所有组分。Luna Cell Ready 裂解模块整合了 DNase I Luna Cell Ready 蛋白酶的功能,仅需 15 分钟即可完成细胞裂解、RNA 释放和基因组 DNA 去除。2 µl 裂解产物(相当于 0.2–4000 个细胞中的 RNA)即可实现 20 µl 体系的 RT-qPCR 反应。

图二:Luna Cell Ready 染料一步法 RT-qPCR 试剂盒可直接对 5-log 梯度稀释细胞的裂解物进行灵敏精准的 RNA 定量

Luna® Cell Ready 一步法 RT-qPCR 试剂盒            货   号                  #E3030S

梯度稀释的 A549 细胞(100,000-10)使用标准反应条件(37°C 裂解 10 分钟,25°C 灭活 5 分钟),在 50 µl Luna Cell Ready 裂解体系(NEBE3032)中裂解。使用 Luna 通用一步法 RT-qPCR 试剂盒(NEBE3005)加入 1 μl 细胞裂解物(相当于 20 μl RT-qPCR 反应体系中 0.2-2,000 个细胞)对目的基因(GOI)进行定量,每个浓度的样品都做了重复。结果展示了两个高频靶标(A):β-actin 和一个低频靶标(B):SMG1。右图展示了扩增效率(E)和线性关系(R2)。

 

图三:Luna Cell Ready 染料一步法 RT-qPCR 试剂盒与提纯 RNA 均可对 5-logs 细胞梯度样品进行精准可信的 RNA 定量

Luna® Cell Ready 一步法 RT-qPCR 试剂盒            货   号                  #E3030S

 

梯度稀释的 A549 细胞(100,000-10)使用标准反应条件(37°C 裂解 10 分钟,25°C 灭活 5 分钟),在 50 µl Luna Cell Ready 裂解体系(NEBE3032)中裂解,也可使用柱提法提纯 RNAA.使用 Luna 通用一步法 RT-qPCR 试剂盒(NEBE3005)对目的基因(GOI)进行定量,加入 1 μl 细胞裂解物(实心圆)或纯化的 RNA(空心圆)(相当于 20 µl RT-qPCR 反应中有 0.2-2,000 个细胞),每个浓度的样品都做了重复。左图:在 5-logs 细胞梯度中检测高频靶标 β-actin和两个低频靶标 ARF3 Tubulin。每个靶标的扩增效率在图中左下方显示。B. A 图中 5-log 细胞梯度稀释样品的目标基因相对于 β-actin Cq 值(ΔCqGOI)),平均 Cqs 通过横线指示。

 

图四:Luna Cell Ready 染料一步法 RT-qPCR 试剂盒的扩增效率及灵敏度均优于其它市售的细胞裂解一步法 RT-qPCR 试剂盒。

 

Luna® Cell Ready 一步法 RT-qPCR 试剂盒            货   号                  #E3030S

梯度稀释的 A549 细胞(100,000-10)使用标准反应条件,在 50 µl Luna Cell Ready 裂解体系(NEBE3032)中裂解。相同的样品使用 Bio-RadSingleShot SYBR Green One-Step Kit, #172-5095)、QiagenSingleShot SYBR Green One-Step Kit, #172-5095)和 Thermo FisherCells-to-CT 1-Step PowerSYBR Green Kit, A25600)并分别按照厂家说明书进行裂解。然后使用每个试剂盒中的一步法 RT-qPCR 模块,加入 1 μl 细胞裂解物(相当于 20 μl RT-qPCR 反应体系中 0.2-2,000 个细胞),对目的基因进行定量,每个浓度的样品都做了重复。对于所有试剂盒,5-log 梯度稀释细胞均显示了 β-actin,(高频靶标)和 RPL32Tubulin (低频靶标)的扩增。为了使结果标准化,将总荧光的 12 设置为阈值。图中显示了扩增效率(E)及最高与最低的 Cq 值。

 

图五:对 24 个靶标测试显示,Luna Cell Ready 染料一步法 RT-qPCR 试剂盒与其它市售的细胞裂解一步法 RT-qPCR 试剂盒相比,灵敏度最高。

Luna® Cell Ready 一步法 RT-qPCR 试剂盒            货   号                  #E3030S

2500 A549 细胞,使用标准反应条件,在 50 µl Luna Cell Ready 裂解体系(NEBE3032)中裂解。或使用市售的 Bio-RadSingleShot SYBR Green One-Step Kit, #172-5095)、QiagenSingleShot SYBR Green One-Step Kit, #172-5095)和 Thermo FisherCells-to-CT 1-Step PowerSYBR Green Kit, A25600)并分别按照厂家说明书进行裂解。每个试剂盒做两个生物重复。然后使用来自每个试剂盒的一步法 RT-qPCR 模块,加入 1 μl 细胞裂解物(相当于 20 µl RT-qPCR 反应中 50 个细胞),对 24 个目标基因进行定量,每个生物样品均做重复。结果显示了 NEB (实心橙色圆圈),BioRad(空心正方形),Qiagen(空心三角形)和 Thermo Fisher(十字形)的平均 Cqs。为了使结果标准化,将总荧光的 12 设置为阈值。Luna Cell Ready 染料一步法 RT-qPCR 试剂盒显示了不同表达水平下 23/24 的基因 Cq 值最早,平均比 Bio-Rad 2.3 Cq,比 Qiagen 3.8 Cq,比 Thermo Fisher 3.6 Cq

 

试剂盒组分

随该产品提供的试剂

 

 

储存(°C

 

浓度

Luna Cell Ready Lysis Buffer

-20

2 X

Luna Cell Ready RNA Protection Reagent

-20

25 X

Luna Cell Ready Protease

-20

25 X

Luna Cell Ready Stop Solution

-20

10 X

DNase I RNase-free

-20

10 X

Luna® WarmStart® RT Enzyme Mix

-20

20 X

Luna® Universal One-Step Reaction Mix

-20

2 X

Nuclease-free Water

-20

 

 

实验需要但不提供的试剂耗材

   磷酸盐缓冲液(PBS

   目标特异性引物

   细胞

   Eppendorf 管,PCR 联管

   PCR

   qPCR 仪器

  移液器和枪头(为减少交叉污染,应使用带滤芯的枪头)

   储存温度

  -20

Luna® Cell Ready 探针一步法 RT-qPCR 试剂盒 货 号 #E3031S-NEB酶试剂 New England Biolabs

上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。

产品资料 – DNA聚合酶与扩增技术 – qPCR & RT-qPCR

Luna® Cell Ready 探针一步法 RT-qPCR 试剂盒                              收藏

货 号
规 格
价 格(元)
北京库存
上海库存
广州库存
成都库存
苏州库存

#E3031S
100 rxns
11,059.00

Download:       

  • isoschizomers     |
  • compatible ends     | 
  • single letter code

特性

 无需纯化,直接从细胞到 RNA 定量

   试剂盒包含 100 次裂解反应和 500 次探针一步法 RT-qPCR 反应

   有效裂解多种细胞系的 10 10 万个细胞

  仅需 15 分钟即可完成细胞裂解、RNA 释放和基因组 DNA 去除

   配合使用 LunaWarmStart® RT Hot Start Taq,提高了热稳定性且可在室温下建立反应体系

   可单独购买裂解模块:Luna Cell Ready 裂解模块(NEBE3032

 

概述

 Luna Cell Ready 探针一步法 RT-qPCR 试剂盒为直接探针法 RNA 检测和定量提供了所有必要的组分,且无需进行 RNA 提取和纯化。Luna Cell Ready 探针一步法 RT-qPCR 试剂盒包含两个模块:1Luna Cell Ready 裂解模块(NEBE3032S)和 2Luna 通用探针一步法 RT-qPCR KitNEBE3006L)。

细胞培养常用于替代活体生物来分析基因表达或对治疗的反应。传统上使用柱提法或化学试剂法从处理过的细胞中提取和纯化 RNA

 

Luna Cell Ready 裂解模块整合了 DNase I Luna Cell Ready 蛋白酶的功能,仅需 15 分钟即可完成细胞裂解、RNA 释放和基因组 DNA 去除。裂解模块包含独特的 Luna Cell Ready RNA 保护试剂,可在细胞裂解过程中保持 RNA 完整性。50 µl 裂解体系下可对 10-10 万个细胞进行裂解反应。2 µl 裂解产物 (相当于 0.2–4000 个细胞的 RNA 即可实现 20 µl 体系的 RT-qPCR 反应。与其它 Luna 产品一样,裂解缓冲液包含惰性蓝色示踪染料,使整个加样流程可视化。 

Luna 通用探针一步法 RT-qPCR 试剂盒可兼容大多数实时荧光定量仪器的各个通道,进行目标 RNA 的实时定量分析。在该试剂盒中,联合使用热启动 Taq DNA 聚合酶与新型 Luna WarmStart 反转录酶,通过适配体可逆抑制来双重控制酶活性。Luna 通用探针一步法 RT-qPCR 试剂盒可以直接将 Luna Cell Ready 裂解模块制备的细胞裂解物中的 RNA 转录本进行检测和定量(单重或多重)。

下表列出的细胞系可使用 Luna Cell Ready 裂解模块裂解。这些细胞进行 5 log 梯度稀释(100000-10 细胞/50 µl 裂解反应),并取 1 ul 裂解产物进行 20 µl 体系的一步法 RT-qPCR 反应。每个细胞系的线性结果区域显示在最后一列。此外,经测试一些昆虫细胞系也能用上述裂解液裂解。

 

 

表一:经验证过的细胞系

 

细胞系

 

特性

菌种

50 ml 裂解体系中的细胞数量

A549

Adherent

H. sapiens, Lung, carcinoma

10 – 100,000

HEK293

Adherent

H. sapiens, Kidney

10 – 100,000

HeLa

Adherent

H. sapiens, Cervix, adenocarcinoma

10 – 100,000

HepG2

Adherent

H. sapiens, Liver, carcinoma

10 – 100,000

NCI-H460

Adherent

H. sapiens, Lung, carcinoma

10 – 100,000

SK-N-SH

Adherent

H. sapiens, Brain, Neuroblastoma

10 – 100,000

U2Os

Adherent

H. sapiens, Bone, osteosarcoma

10 – 10,000

Jurkat

Suspension

H. sapiens, T lymphocyte, leukemia

10 – 100,000

K-562

Suspension

H. sapiens, Lymphoblast, leukemia

10 – 1,000

 图一:Luna Cell Ready 一步法 RT-qPCR 实验流程

 Luna® Cell Ready 探针一步法 RT-qPCR 试剂盒            货   号                  #E3031S

Luna Cell Ready 一步法 RT-qPCR 试剂盒提供了直接从细胞(50 µl 体系可裂解高达 10 万个细胞)到 RNA 检测和定量的所有组分。Luna Cell Ready 裂解模块整合了 DNase I Luna Cell Ready 蛋白酶的功能,仅需 15 分钟即可完成细胞裂解、RNA 释放和基因组 DNA 去除。2 µl 裂解产物(相当于 0.2–4000 个细胞中的 RNA)即可实现 20 µl 体系的 RT-qPCR 反应。 

 

图二:Luna Cell Ready 探针一步法 RT-qPCR 试剂盒可直接对 5-log 梯度稀释细胞的裂解物进行灵敏精准的 RNA 定量

 

Luna® Cell Ready 探针一步法 RT-qPCR 试剂盒            货   号                  #E3031S

梯度稀释的 A549 细胞(100,000-10)使用标准反应条件(37°C 裂解 10 分钟,25°C 灭活 5 分钟),在 50 µl Luna Cell Ready 裂解体系(NEBE3032)中裂解。使用 Luna 通用探针一步法 RT-qPCR 试剂盒(NEBE3006),加入 1 μl 细胞裂解物(相当于 20 μl RT-qPCR 反应体系中 0.2-2,000 个细胞)对目的基因进行定量,每个浓度的样品都做了重复。结果展示了两个高频靶标:β-actinTexas Red)和 GAPDHFAM),一个低频靶标:RPL32HEX),的多重结果(A)和单重结果(B),以及相应的扩增效率(E)和线性关系(R2)。(C)所有三个目标的多重和单重的 Cq 叠加证明了这些结果的一致性。

三:Luna Cell Ready 探针一步法 RT-qPCR 试剂盒与提纯 RNA 均可对 5-logs 细胞梯度样品进行精准可信的 RNA 定量

 Luna® Cell Ready 探针一步法 RT-qPCR 试剂盒            货   号                  #E3031S

梯度稀释的 A549 细胞(100,000-10)使用标准反应条件(37°C 裂解 10 分钟,25°C 灭活 5 分钟),在 50 µl Luna Cell Ready 裂解体系(NEBE3032)中裂解,也可使用柱提法提纯 RNAA.使用 Luna 通用探针一步法 RT-qPCR 试剂盒(NEBE3006)对目的基因进行定量,加入 1 μl 细胞裂解物(实心圆,左)或纯化的 RNA(空心圆,右)(上样量相当于 20 µl RT-qPCR 反应中有 0.2-2,000 个细胞),每个浓度的样品都做了重复。在 5-logs 细胞梯度中检测高频靶标 β-actin 和两个低频靶标 ARF3 Tubulin。每个靶标的扩增效率在图中左下方显示。

图四:Luna Cell Ready 染料一步法 RT-qPCR 试剂盒的扩增效率及灵敏度均优于其它市售的细胞裂解一步法 RT-qPCR 试剂盒。

Luna® Cell Ready 探针一步法 RT-qPCR 试剂盒            货   号                  #E3031S

梯度稀释的 A549 细胞(100,000-10)使用标准反应条件,在 50 µl Luna Cell Ready 裂解体系(NEBE3032)中裂解。相同的样品使用 Bio-RadSingleShot™ Probes One-Step Kit for Cell Lysis and RT-qPCR, #172-5070)、QiagenFastLane Cell Probe Kit, #216413)和 Thermo FisherCells-to-CT 1-Step TaqMan Kit, A25603并分别按照厂家说明书进行裂解。然后使用每个试剂盒中的一步法 RT-qPCR 模块,加入 1 μl 细胞裂解物(相当于 20 μl RT-qPCR 反应体系中 0.2-2,000 个细胞),对目的基因进行定量,每个浓度的样品都做了重复。5-log 梯度稀释的细胞通过多重实验测试了 β-actin Tye)、GAPDH FAM)和 RPL32 Cy5的扩增。为了使结果标准化,将每个试剂盒总荧光值的 10%(Tye)、5%(FAM)、15%(Cy5)设置为阈值。图中显示了扩增效率(E)及最高与最低的 Cq 值。

试剂盒组分

随该产品提供的试剂

 

储存(°C

浓度

Luna Cell Ready Lysis Buffer

-20

2 X

Luna Cell Ready RNA Protection Reagent

-20

25 X

Luna Cell Ready Protease

-20

25 X

Luna Cell Ready Stop Solution

-20

10 X

DNase I RNase-free

-20

10 X

Luna® WarmStart® RT Enzyme Mix

-20

20 X

Luna® Universal Probe One-Step Reaction Mix

-20

2 X

Nuclease-free Water

-20

 

 实验需要但不提供的试剂耗材

  磷酸盐缓冲液(PBS

   目标特异性引物

   细胞

   Eppendorf

   PCR联管

   PCR

   qPCR 仪器

   移液器和枪头(为减少交叉污染,应使用带滤芯的枪头)

 储存温度

-20

Luna® Cell Ready 裂解模块 货 号 #E3032S-NEB酶试剂 New England Biolabs

上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。

产品资料 – DNA聚合酶与扩增技术 – qPCR & RT-qPCR

Luna® Cell Ready 裂解模块                              收藏

货 号
规 格
价 格(元)
北京库存
上海库存
广州库存
成都库存
苏州库存

#E3032S
100 rxns
5,959.00

Download:       

  • isoschizomers     |
  • compatible ends     | 
  • single letter code

特性

 ·仅需 15 分钟即可完成细胞裂解、RNA 释放和基因组 DNA 去除
 ·一次反应高效可裂解 10 至 10 万个细胞,适用于众多细胞系
 ·与其它 RNA 提取方法相比减少了样品损失,尤其对于低细胞起始量。
 ·兼容高通量应用,可在室温下进行细胞裂解
 ·与 Luna 通用一步法 RT-qPCR 试剂盒搭配使用时性能最佳:NEB#E3005(染料法)或 NEB#E3006(探针法) 
 ·提供完整的细胞裂解和一步法 RT-qPCR 试剂盒方案:NEB #E3030(染料法)或 NEB #E3031(探针法)

概述

   细胞培养常用于替代活体生物来分析基因表达或对治疗的反应。传统上使用柱提法或化学试剂法从处理过的细胞中提取和纯化 RNA.Luna Cell Ready 裂解模块无需进行传统的 RNA 提取步骤即可评估RNA表达水平,是一种快速、方便且灵敏的替代方案。该模块可同时进行细胞裂解、RNA 释放、基因组 DNA 去除,获得的细胞裂解物可直接使用Luna通用一步法 RT-qPCR 试剂盒进行 RT-qPCR 分析。与其它 Luna 产品一样,裂解缓冲液包含惰性蓝色示踪染料,使整个加样流程可视化。

下表列出的细胞系可使用 Luna Cell Ready 裂解模块裂解。这些细胞进行 5 log 梯度稀释(100000-10 细胞/50 µl 裂解反应),并取 1 ul 裂解产物进行 20 µl 体系的一步法 RT-qPCR 反应。每个细胞系的线性结果区域显示在最后一列。此外,经测试一些昆虫细胞系也能用上述裂解液裂解。

 

表一:经验证过的细胞系

细胞系

特性

菌种

50 ml 裂解体系中的细胞数量

A549

Adherent

H. sapiens, Lung, carcinoma

10 – 100,000

HEK293

Adherent

H. sapiens, Kidney

10 – 100,000

HeLa

Adherent

H. sapiens, Cervix, adenocarcinoma

10 – 100,000

HepG2

Adherent

H. sapiens, Liver, carcinoma

10 – 100,000

NCI-H460

Adherent

H. sapiens, Lung, carcinoma

10 – 100,000

SK-N-SH

Adherent

H. sapiens, Brain, Neuroblastoma

10 – 100,000

U2Os

Adherent

H. sapiens, Bone, osteosarcoma

10 – 10,000

Jurkat

Suspension

H. sapiens, T lymphocyte, leukemia

10 – 100,000

K-562

Suspension

H. sapiens, Lymphoblast, leukemia

10 – 1,000

 

图一:Luna Cell Ready 一步法 RT-qPCR 实验流程

 

Luna® Cell Ready 裂解模块            货   号                  #E3032S

 

Luna Cell Ready 裂解模块整合了 DNase I 和 Luna Cell Ready 蛋白酶的功能,可简单高效的进行细胞裂解,仅需 15 分钟即可完成细胞裂解、RNA 释放和基因组 DNA 去除。2 µl 裂解产物(相当于 0.2–4000 个细胞中的 RNA)即可实现 20 µl 体系的 RT-qPCR 反应。

 

试剂盒组分

 

随该产品提供的试剂

 

储存(°C

浓度

Luna Cell Ready Lysis Buffer

-20

2 X

Luna Cell Ready RNA Protection Reagent

-20

25 X

Luna Cell Ready Protease

-20

25 X

Luna Cell Ready Stop Solution

-20

10 X

DNase I RNase-free

-20

10 X

 实验需要但不提供的试剂耗材

  •    磷酸盐缓冲液(PBS
  •    目标特异性引物
  •    细胞
  •    Eppendorf
  •    PCR 联管
  •    PCR
  •    移液器和枪头(为减少交叉污染,应使用带滤芯的枪头)

    储存温度

 -20℃

Engrafted Cell Markers酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Engrafted Cell Markers
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y40400 STEM101 50 μg ¥1,958 Engrafted Cell Markers Engrafted Cell Markers Engrafted Cell Markers
Cellartis Y40410 STEM121 50 μg ¥1,958 Engrafted Cell Markers Engrafted Cell Markers Engrafted Cell Markers
Cellartis Y40420 STEM123 50 μg ¥1,958 Engrafted Cell Markers Engrafted Cell Markers Engrafted Cell Markers
收藏产品 加入购物车
监控人类细胞移植用途的抗体
(Antibodies for Detecting Human Cells Transplanted into Mice and Rats)
STEM101、STEM121、STEM123是三种小鼠来源的单克隆抗体,用于人类细胞向小鼠和大鼠移植实验中,监控细胞移植、转移、分化。
 
■ 产品特点
· STEM101 reacts specifically with the nuclear human Ku80 protein; the antibody does not cross-react with mouse or rat Ku80 in brain tissue or cell extracts
Note: STEM101 (Y40400) is not suitable for immunohistochemically staining on paraffin embedded tissue sections. We recommend using fresh frozen tissue with this antibody.
· STEM121 reacts with a human cytoplasmic protein expressed in brain, liver, pancreas, and CNS cells the antibody does not cross-react with brain tissue or cell extracts from mouse, rat, or cynomologous monkey
· STEM123 reacts specifically with human GFAP, a marker of astroglial cells; the antibody does not cross-react with brain tissue or cell extracts from mouse or rat
 
■ 产品应用
· Localization, quantification, and/or characterization of engrafted cells of human origin by immunohistochemistry and immunofluorescence
 
 
 
■ 产品详情请点击:Engrafted Cell Markers
 
Engrafted Cell Markers
STEM101 detects nuclei of transplanted human neural stem cells in the olfactory bulb of a mouse brain.
 
Engrafted Cell Markers
STEM121 detects migration and differentiation of transplanted human neural stem cells in the hippocampus of a mouse brain.
 
Engrafted Cell Markers
STEM123 detects human GFAP+ astrocytes differentiated from human neural stem cells after transplantation into a mouse brain.
 
参考文献:
1. Cummings BJ, et al. (2005) Human neural stem cells differentiate and promote locomotor recovery in spinal cord-injured mice. PNAS 102: 14069-14074.
2. Guzman R, et al. (2007) Long-term monitoring of transplanted human neural stem cells in developmental and pathological contexts with MRI. PNAS 104: 10211-10216.
3. Tamaki SJ, et al. (2009) Neuroprotection of Host Cells by Human Central Nervous System Stem Cells in a Mouse Model of Infantile Neuronal Ceroid Lipofuscinosis. Cell Stem Cell 5:310-319.
4. Salazar DL, et al. (2010) Human Neural Stem Cells Differentiate and Promote Locomotor Recovery in an Early Chronic Spinal Cord Injury NOD-scid Mouse Model. PLoS ONE 5: e12272.
5. Kelly S, et al. Transplanted human fetal neural stem cells survive, migrate, and differentiate in ischemic rat cerebral cortex. PNAS (2004) 101:11839 11844.
6. Cummings BJ, et al. Human neural stem cells differentiate and promote locomotor recovery in spinal cord-injured mice. PNAS (2005) 102: 14069-14074.
7. Tamaki SJ, et al. Neuroprotection of Host Cells by Human Central Nervous System Stem Cells in a Mouse Model of Infantile Neuronal Ceroid Lipofuscinosis. Cell Stem Cell (2009) 5:310-319.
8. Kallur T, et al. Human Fetal Cortical and Striatal Neural Stem Cells Generate Region-Specific Neurons In Vitro and Differentiate Extensively to Neurons After Intrastriatal Transplantation in Neonatal Rats. J Neurosci Res. (2006) 84:1630-1644.
9. Salazar DL, et al. Human Neural Stem Cells Differentiate and Promote Locomotor Recovery in an Early Chronic Spinal Cord Injury NOD-scid Mouse Model. PLoS ONE (2010) 5: e12272.
10. Tamaki SJ, et al. (2009) Neuroprotection of Host Cells by Human Central Nervous System Stem Cells in a Mouse Model of Infantile Neuronal Ceroid Lipofuscinosis. Cell Stem Cell 5:310-319.
11. Salazar DL, et al. (2010) Human Neural Stem Cells Differentiate and Promote Locomotor Recovery in an Early Chronic Spinal Cord Injury NOD-scid Mouse Model. PLoS ONE 5:e12272.
12. Cummings BJ, et al. (2005) Human neural stem cells differentiate and promote locomotor recovery in spinal cord-injured mice. PNAS 102: 14069-14074.
 

Human Stem Cell Culture System (DEF-CS, all-in-one format)酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Human Stem Cell Culture System (DEF-CS, all-in-one format)
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y30010 Cellartis® DEF-CS 500 Culture System 1 Kit ¥5,979 Human Stem Cell Culture System (DEF-CS, all-in-one format) Human Stem Cell Culture System (DEF-CS, all-in-one format) Human Stem Cell Culture System (DEF-CS, all-in-one format)
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Human iPS和Human ES增殖用培养基
(DEF-CS Culture System, all-in-one format)
作为一款高端创新型的产品,Cellartis DEF-CS培养系统使得单细胞干细胞操作成为常规的操作。该产品是专门针对人类诱导干细胞(hiPS)和人类胚胎干细胞(hES)研发的高效率增殖用培养基产品体系。该产品系统为all-in-one型,包括了所有组分,客户不需要另外求购组分。该产品是成分确定的培养基产品,而且不需要饲养层。DEF-CS既可以实现单细胞培养,也可以用于传统的iPS培养模式以及大规模干细胞增殖。在使用DEF-CS培养基增殖干细胞时,几乎没有背景分化的问题,这使得细胞筛选不再必需。作为一款创新产品,利用DEF-CS培养干细胞时,可以使用酶消化法(enzymatic passaging)实现需要单细胞操作,这一特点十分有利于高通量细胞鉴别筛选(high-throughput screening)、转染(transfection)、框架接种(scaffold seeding)等。
 
■ 产品特点
· Feeder-free and chemically defined culture system for efficient expansion of human iPS cells
· Ideal system for culturing iPS cells; cells maintain pluripotency and long-term potential for self-renewal and differentiation
· Complete kit includes 500 ml basal medium, additives, and coating compound
· Robust system provides high reproducibility and stable growth
· Maintains cells in an undifferentiated state with virtually no background differentiation
 
■ 产品应用
· Scale-up and mass production of human iPS cells
· Single-cell culture of human iPS cells
· Transfection and reprogramming
· High-throughput screening
· Tissue engineering (seeding cells on a scaffold)
 
■ 产品详情请点击:Human Stem Cell Culture System (DEF-CS, all-in-one format)
 
Human Stem Cell Culture System (DEF-CS, all-in-one format)
Robust growth of human induced pluripotent stem (iPS) cells in the Cellartis DEF-CS Culture System. The number of iPS cells was quantified after being cultured for three weeks using either the Cellartis DEF-CS Culture System, a reference feeder system, or four other stem cell culture systems.
 
参考文献:
1. Sivertsson, Louise, et al. "Hepatic differentiation and maturation of human embryonic stem cells cultured in a perfused three-dimensional bioreactor." Stem cells and development 22.4 (2012): 581-594.
2. Hanson, Charles, et al. "Transplantation of human embryonic stem cells onto a partially wounded human cornea in vitro." Acta ophthalmologica 91.2 (2013): 127-130.
3. Norrman, Karin, et al. "Distinct gene expression signatures in human embryonic stem cells differentiated towards definitive endoderm at single-cell level." Methods 59.1 (2013): 59-70.
4. Ulvestad, Maria, et al. "Drug metabolizing enzyme and transporter protein profiles of hepatocytes derived from human embryonic and induced pluripotent stem cells." Biochemical pharmacology 86.5 (2013): 691-702.
5. Ramirez JM, et al. Side scatter intensity is highly heterogeneous in undifferentiated pluripotent stem cells and predicts clonogenic self-renewal. Stem Cells Dev.2013 Jun 15;22(12):1851-1860.
6. Borestrom, Cecilia, et al. “Footprint-free human induced pluripotent stem cells from articular cartilage with redifferentiation capacity: A first step toward a clinical-grade cell source.” Stem Cells Trans. Med. (2014) 3, 433-447.
7. Kia, Richard, et al. "MicroRNA-122: a novel hepatocyte-enriched in vitro marker of drug-induced cellular toxicity." Toxicological Sciences (2014): kfu269.
8. Valton, Julien, et al. "Efficient strategies for TALEN-mediated genome editing in mammalian cell lines."  Methods 69.2 (2014): 151-170.
9. Zandén, Carl, et al. "Stem cell responses to plasma surface modified electrospun polyurethane scaffolds." Nanomedicine: Nanotechnology, Biology and Medicine 10.5 (2014): 949-958.
10. Asplund, Annika, et al. “One Standardized Differentiation Procedure Robustly Generates Homogenous Hepatocyte Cultures Displaying? Metabolic Diversity from a Large Panel of Human Pluripotent Stem Cells” Stem Cell Rev and Rep (2015)
 
 
 
 

iPS Cell to Hepatocyte Differentiation System酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

iPS Cell to Hepatocyte Differentiation System
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y30055 Cellartis® iPS Cell to Hepatocyte Differentiation System 1 Kit ¥13,846 iPS Cell to Hepatocyte Differentiation System iPS Cell to Hepatocyte Differentiation System iPS Cell to Hepatocyte Differentiation System
Cellartis Y30051 Cellartis® Hepatocyte Maintenance Medium 100 ml ¥2,198 iPS Cell to Hepatocyte Differentiation System iPS Cell to Hepatocyte Differentiation System iPS Cell to Hepatocyte Differentiation System
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iPS向肝脏细胞定向分化操作系统
(Cellartis iPS Cell to Hepatocyte Differentiation System)
以肝脏细胞为模型的实验越来越受到重视,在肝脏病变机理研究、药物代谢研究、药物毒理评估等方面有广阔的应用前景。然而,以个体原代肝细胞为材料来源会产生如取材困难、材料批间差大、无法长期稳定供应等一系列难以克服的问题。因此,通过iPS批量转化生产肝细胞的方法就显示出特别的优势。Cellartis iPS Cell to Hepatocyte Differentiation System可以稳定地、规模化地实现iPS向hepatocyte的诱导转化生产,而且诱导转化出的肝细胞能够稳定表达药物代谢相关酶系统和药物转运系统。
Cellartis iPS Cell to Hepatocyte Differentiation System是一套Do-It-Yourself系统,客户既可以对某个特殊病人来源的iPS实现转化,也可以利用cellartis供应的iPS进行标准化操作。使用本产品的过程起始于诱导人iPS转化成为确定性内胚层(Definitive Endoderm (DE))细胞,然后诱导转化DE至肝细胞。本产品是All-In-One型产品,实验所需的培养基、基质(coating)等包括在内。应用本产品转化约3 × 106 hiPS cells,可以获得约5 × 106 肝细胞(大约50 cm2面积的单层贴壁细胞)。
利用Cellartis iPS Cell to Hepatocyte Differentiation System转化得到的肝细胞除了清晰表现出肝细胞的必备特征外(如肝细胞特异性标志物、CYP系列酶等),还保留了原始细胞供体者的遗传基因背景。另外,相比较从组织切除获得的原代肝脏细胞(通常需要低温保存),通过本产品系统转化获得的肝细胞在更长的时间跨度上保持着生理功能的稳定性。这一点对于毒理学评估和病毒感染评估具有显著的意义。为长时间维持培养hiPS转化而来的肝细胞,我们推荐Cellartis Hepatocyte Maintenance Medium (Cat. No. Y30051)。
 
关于Cellartis 卓越的iPS Cell to Hepatocyte Differentiation技术的专项说明,请点击:
详情
 
■ 产品特点
· Highly reproducible, robust system—the exact same protocol has been shown to work across 25 different iPS cell lines, so no need to optimize for your lines.
· Ideal for drug metabolism and safety studies—consistently generate panels of >90% pure, functional, hiPS cell-derived hepatocytes with diverse genetic backgrounds.
· Customized starting materials—start with any disease-relevant iPS cell lines and create accurate liver disease models.
· Ready for personalized medicine—make patient-specific, disease-specific cells for therapeutic applications.
· Extended experimental window—hepatocytes can be used for a minimum of 11 days for functional tests.
 
■ 产品成分
· Cellartis DEF-CS 100 Culture System (Y30020, not sold separately)
· Cellartis Definitive Endoderm Differentiation Kit (Y30030, not sold separately)
· Cellartis Hepatocyte Differentiation Kit (Y30050)
 
■ 产品详情请点击:iPS Cell to Hepatocyte Differentiation System
 
iPS Cell to Hepatocyte Differentiation System
Immunocytochemistry analysis of hepatocyte differentiation. hiPS cells were differentiated into functional hepatocytes using the Cellartis iPS Cell to Hepatocyte Differentiation System. Hepatocytes were immunostained to detect early hepatic markers HNF4α (red, nuclear) and CK18 (green) on days 14 and 21. As the hepatocytes matured, expression of liver-specific markers CYP3A (red, cytoplasmic) and Albumin (green) increased as seen on day 28, as expected. Cell nuclei were stained with DAPI (blue).
 
参考文献:
Asplund, Annika, et al.One Standardized Differentiation Procedure Robustly Generates Homogenous Hepatocyte Cultures Displaying? Metabolic Diversity from a Large Panel of Human Pluripotent Stem Cells. Stem Cell Rev and Rep (2015).
 
用户分享:
◆ 东海大学医学部-纸谷聪英教授
   利用人iPS细胞体外构建多囊性肝病(Polycystic liver disease)模型
 
iPS Cell to Hepatocyte Differentiation System
纸谷聪英教授使用Cellartis iPS Cell to Hepatocyte Differentiation System(Y30055)构建肝脏疾病模型的研究成果已刊登在Stem Cell Research。
【论文】
Akihide, K., Hiromi, C., Kinuyo, I., Emi, A., Kota, T., & Tatehiro, K., et al. (2018). An in vitro model of polycystic liver disease using genome-edited human inducible pluripotent stem cells. Stem Cell Research, 32, 17-24.
 
实验数据
 
iPS Cell to Hepatocyte Differentiation System
 
图:使用 Cellartis iPS Cell to Hepatocyte Differentiation System(Y30055)获得的人iPSC来源肝前体细胞(iPS-HPCs)诱导分化为胆管状Cyst结构。
(引用自文献Akihide Kamiya et al., Stem Cell Research, 2018)
 
访谈时间:
1. 开始使用Cellartis iPS Cell to Hepatocyte Differentiation System(Y30055)的契机是什么?
很多研究组已经报道了多种从人iPS细胞诱导分化形成肝脏细胞的方法,我们研究组也在做相关工作。由于细胞分化不稳定,我担心使用饲养层细胞培养人iPS细胞会对肝脏分化产生影响,因此决定尝试使用Cellartis iPS细胞无饲养层培养系统Cellartis DEF-CS™ 500 Culture System(Y30010)和肝脏分化系统Cellartis iPS Cell to Hepatocyte Differentiation System(Y30055)。
 
2.实际使用后感觉如何?
与之前使用的饲养层培养方法相比,Y30010能更简便高效的进行培养;使用Y30055进行的肝脏分化非常稳定,进一步提升了实验效率。
 
3. 对暂未使用该制品的用户有什么寄语?
人iPS细胞向肝脏细胞分化过程中,细胞密度对分化效率有很大影响,进而影响实验结果。我认为Cellartis系统的优点主要在于各个实验阶段偏差小,结果稳定,适合追求实验效率的人。
 
 
◆ 东京医科齿科大学-柿沼晴教授
   先天性肝纤维化模型的开发和病理阐明的研究
 
iPS Cell to Hepatocyte Differentiation System
柿沼晴教授使用Cellartis iPS Cell to Hepatocyte Differentiation System(Y30055)构建肝脏疾病模型的研究成果已刊登在Journal of Hepatology。
 
【论文】
Tsunoda, T., Kakinuma, S., Miyoshi, M., Kamiya, A., Kaneko, S., & Sato, A., et al. (2019). Loss of fibrocystin promotes interleukin-8-dependent proliferation and ctgf production of biliary epithelium. Journal of Hepatology.
 
访谈时间:
1.开始使用Cellartis iPS Cell to Hepatocyte Differentiation System(Y30055)的契机是什么?
一起进行研究的老师使用后能够进行稳定的培养,我们研究室也正好想利用无饲养层培养系统,所以开始使用了Cellartis DEF-CS™ 500 Culture System(Y30010)和肝脏分化系统Cellartis iPS Cell to Hepatocyte Differentiation System(Y30055)。
 
2.实际使用后感觉如何?
Y30010无饲养层培养系统使用比较简便,同时能够稳定的培养人iPS细胞。这次实验中,利用Y30055分化至肝前体细胞(iPS-HPCs),进一步诱导分化成了胆管细胞,我认为诱导分化的再现性好。不仅分化至成熟肝细胞谱系,同时相比于以往报道的方法,再现性更稳定。
 
3. 对暂未使用该制品的用户有什么寄语?
在人iPS细胞的培养和分化至肝细胞谱系的诱导中,即使是对人iPS细胞使用经验尚浅的实验者,也能够进行培养并得到稳定的再现。
 
 

Human Stem Cell Derived Hepatocytes酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Human Stem Cell Derived Hepatocytes
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y10133 Cellartis® Enhanced hiPS-HEP v2 (from ChiPSC12) Kit 1 kit ¥21,173 Human Stem Cell Derived Hepatocytes Human Stem Cell Derived Hepatocytes Human Stem Cell Derived Hepatocytes
Cellartis Y10134 Cellartis® Enhanced hiPS-HEP v2 (from ChiPSC18) Kit 1 kit ¥21,173 Human Stem Cell Derived Hepatocytes Human Stem Cell Derived Hepatocytes Human Stem Cell Derived Hepatocytes
Cellartis Y10135 Cellartis® Enhanced hiPS-HEP v2 (from ChiPSC22) Kit 1 kit ¥21,173 Human Stem Cell Derived Hepatocytes Human Stem Cell Derived Hepatocytes Human Stem Cell Derived Hepatocytes
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无标题文档

人iPS干细胞分化来的肝细胞
Cellartis enhanced hiPS-HEP v2 kits 提供了可持续供应的、成熟的人诱导多能干细胞(hiPS)来源的肝脏细胞。在超过14天的检测窗口期,hiPS-HEP细胞拥有功能性的药物代谢机制,使其成为体外药物研发、药物代谢、和相关毒性研究的理想选择;具备成熟、稳健的代谢特征,支持其作为代谢疾病模型。方便使用的试剂盒包括完全的培养基和组份,用于长期检测。与其他肝脏细胞模型相比,培养的Cellartis hiPS-HEP细胞存活时间更长,进而拥有更长的检测窗口用于慢性-毒性实验和作为代谢疾病模型研究。
 
■ 产品特点
· Complete kit for thawing, plating, and culturing the provided cells
· Highly homogeneous population with consistent performance between batches and over time
· Cells are functional and stable for a 14-day assay window
· Cells show long-term, stable CYP450 activities
· Cells express key hepatocyte proteins, including α1AT, Alb, CK18, HNF4α, and E-cadherin
· Cells secrete physiologically relevant levels of albumin and urea
· Cells demonstrate functional insulin and glucose regulation
· Cells show phase I and II enzyme activities
· Cells demonstrate LDL uptake
 
■ 产品组成
· 1 vial of ~1.2 × 107 cells
· Hepatocyte thawing medium
· Hepatocyte coating
· Hepatocyte plating medium
· Hepatocyte washing medium
· Hepatocyte long-term maintenance medium
 
■ 产品应用
· Toxicity testing, including chronic toxicity assays
· Drug discovery and development
· Metabolic studies
· Metabolic disease modeling
 
■ 产品详情请点击:Human Stem Cell Derived Hepatocytes
 
Human Stem Cell Derived Hepatocytes
 
Cellartis 人iPS细胞来源的肝脏细胞可以作为长期的、可靠的肝脏疾病模型。
Cellartis human induced pluripotent stem (iPS) cell-derived hepatocytes are a long-lasting, reliable liver disease model. Ideal for long-term studies, these mature, functional, pure hepatocytes allow you to generate consistent results with low batch-to-batch variability. Cellartis iPSC-derived hepatocytes can be used for extended culture significantly longer than human primary hepatocytes—allowing you to get more data from your chronic toxicity studies.
 
Human Stem Cell Derived Hepatocytes
 
Cellartis enhanced hiPS-HEP cells可以稳定表达CYP450 活性超过21天。
CYP450 activity is stable in enhanced hiPS-HEP cells over 21 days. LC/MS was used to analyze CYP450 activity in cultured enhanced hiPS-HEP cells previously derived from the hiPS cell lines ChiPSC12, ChiPSC18, and ChiPSC22 (abbreviated as C12, C18, and C22). CYP3A (Panel A), CYP2C9 (Panel B), CYP1A (Panel C), and CYP2C19 (Panel D) activities in enhanced hiPS-HEP cells are stable over an extended culture time. Cryopreserved human primary hepatocytes (hphep), which are functional in culture for a significantly shorter time than enhanced hiPS-HEP cells, were thawed and cultured for 20 hours, and their data are shown as the black bar in each panel.
 
Human Stem Cell Derived Hepatocytes
 
Cellartis enhanced hiPS-HEP细胞在20天的培养中表达白蛋白(Albumin)。
Albumin is present in enhanced hiPS-HEP cells through 20 days in culture. Panel A. Representative images of enhanced hiPS-HEP cells from human iPS cell line ChiPSC18 (C18) taken 12 days after thawing (right), compared to cryopreserved human primary hepatocytes (hphep) taken 24 hr after thawing (left). Cells were stained for albumin and DAPI. Panel B. Albumin secretion as measured by ELISA; n=2 for enhanced hiPS-HEP cells, with the exception of C18 at 20 days (n=1), and n=3 donors for hphep.
 
(以上图片均来源于Takara Bio USA, Inc.)
 
参考文献:
1. Heins et al. Stem Cells 2004; 22: 367-376.United States National Stem Cell Bank; http://www.nationalstemcellbank.org.
2. Mantel N et al. Potential markers of attenuation of YF virus after infection of stem cell-derived human hepatocytes with wild-type Asibi or live-attenuated YF17D virus.Supplement to The American Journal of tropical Medicine and Hygiene, Volume 83, November 2010, Number 5, abstract 12.
3. Yildirimman R et al. Human embryonic stem cell derived hepatocyte-like cells as a tool for in vitro hazard assessment of chemical carcinogenicity. Toxicol. Sci. 2011 Dec; 124(2): 278-90.
4. Ulvestad M et al. Drug metabolizing enzyme and transporter protein profi les of hepatocytes derived from human embryonic and induced pluripotent stem cells. Biochem Pharmacol. 2013 Sep 1; 86(5):691-702.
 
 

Human iPS Cell Generation™ All-in-One Vector酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Human iPS Cell Generation All-in-One Vector
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Takara 3671 Human iPS Cell Generation All-in-One Vector 10 μg ¥10,014 Human iPS Cell Generation™ All-in-One Vector Human iPS Cell Generation™ All-in-One Vector Human iPS Cell Generation™ All-in-One Vector
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□ 浓度 0.5 μg/μl
     
□ 贮存溶液  
     Tris-HCl (pH 8.0) 10 mM
     EDTA 1 mM
Limited Use Label License: [L44], [M71]
 
■ 制品说明
本制品是制备Human iPS细胞诱导用逆转录病毒所使用的载体。在高效、高表达逆转录病毒载体pDON-5 DNA(Code No.: 3658)上,插入了iPS细胞诱导用基因OCT3/4SOX2KLF4LIN28NANOG,各基因以Thosea asigna virus 的2A*1序列连接在一个载体上。本载体与Retrovirus Packaging Kit Ampho(Code No.: 6161)中的pGP Vector以及pE-ampho Vector共转染至逆转录病毒制备用细胞G3T-hi(Code No.: 6163)后,可以制备iPS诱导用逆转录病毒。如此制备的重组逆转录病毒由于和RetroNectin®(Recombinant Human Fibronectin Fragment)(Code No.: T100A/B)有高亲和性,所以在向目的细胞中导入基因时,使用经RetroNectin®处理后的培养皿或平板,可以得到较高的基因导入率,有效提高iPS细胞的诱导效率。
*1 5种基因转录在同一个mRNA上,翻译时在2A序列的特定位点被分割,使5种蛋白质可以等量表达。
 
■ 保存
-20
 
■ 各载体的遗传信息
  基因名   GenBank Accession No.
  OCT4   NM_002701.4
  SOX2   NM_003106.2
  KLF4   NM_004235.3*2
  LIN28   NM_024674.4
  NANOG   NM_024865.2
*2 更新的NM_004235.4中,CDS的5’端追加了对应9个氨基酸的碱基。本制品的KLF4基因中不含N端的这9个氨基酸的序列,已经确认可以有效诱导iPS细胞。
 
■ 载体图谱
Human iPS Cell Generation™ All-in-One Vector
 
 
 
 

Human Stem Cell Derived Intestinal Epithelial Cells酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Human Stem Cell Derived Intestinal Epithelial Cells
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y50035 Cellartis® Intestinal Epithelial Cells (from ChiPSC18) Kit 1 Kit ¥20,754 Human Stem Cell Derived Intestinal Epithelial Cells Human Stem Cell Derived Intestinal Epithelial Cells Human Stem Cell Derived Intestinal Epithelial Cells
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来源于人类iPS细胞的小肠上皮细胞
(Intestinal Epithelial Cells Derived from Human Induced Pluripotent Stem Cells)
Cellartis Intestinal Epithelial Cells (from ChiPSC18) 是来源于人iPS细胞的小肠上皮细胞,具备肠上皮样特征,包括形成紧密连接、表达绒毛蛋白villin和肠转录因子CDX2。与人结肠癌Caco-2细胞系相比,该细胞具有更高的代谢酶CYP3A4和转运蛋白PEPT1的表达,为药物代谢吸收、药物相互作用评估提供更好的预测结果。
Cellartis Intestinal Epithelial Cells (from ChiPSC18) Kit 包括1管冻存的细胞Cellartis Intestinal Epithelial Cells (from ChiPSC18)和1个试剂盒Cellartis IEC Maturation Kit。解冻的Cellartis Intestinal Epithelial Cells (from ChiPSC18)使用Cellartis IEC Maturation Kit预先培养5天后,可用于各种检测。
 
■ 产品特点
· 解冻5天内细胞形成紧密连接,解冻5-8天后可用于检测
· 细胞高表达肠上皮标志物 (villin、CDX2等)
· 与Caco-2细胞相比,拥有更高的代谢酶CYP3A4和转运蛋白PEPT1的表达
· 细胞形成一个功能性的、可渗透的屏障,可用于预测药物在体内的渗透性和吸收
 
■ 产品应用
· 药物在小肠的吸收和代谢评价
· 药物研发中的ADME分析
· 药物渗透性检测
 
■ 产品组成
· 1 vial of~4.8 x 106 cells
· 1 Cellartis IEC Maturation Kit(IEC Maturation Basal Medium;IEC Supplement A;IEC Supplement B;IEC Supplement C;IEC Supplement D)
 
Human Stem Cell Derived Intestinal Epithelial Cells
 
实验例
Human Stem Cell Derived Intestinal Epithelial Cells
图1. 紧密连接蛋白ZO-1的免疫荧光染色结果显示,解冻后第5天,hiPSC-derived intestinal epithelial cells (IEC)细胞间形成紧密连接(如闭锁小带zonula occludens)。
 
Human Stem Cell Derived Intestinal Epithelial Cells
图2. 在解冻后第7天,与Caco-2细胞相比,hiPSC-derived intestinal epithelial cells (IEC)中高表达肠上皮标志物villin和CDX2。
 
Human Stem Cell Derived Intestinal Epithelial Cells
图3. 在解冻后第7天,与Caco-2细胞相比,hiPSC-derived intestinal epithelial cells (IEC)中高表达药物代谢酶CYP3A4和药物转运蛋白PEPT1。
 
 
■ 产品详情请点击:Human Stem Cell Derived Intestinal Epithelial Cells
 
 

Human Stem Cell Derived Beta Cells酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Human Stem Cell Derived Beta Cells
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y10100 Cellartis® hiPS Beta Cells (from ChiPSC12) Kit 1 Kit ¥18,655 Human Stem Cell Derived Beta Cells Human Stem Cell Derived Beta Cells Human Stem Cell Derived Beta Cells
Cellartis Y10106 Cellartis® hiPS Beta Cells (from ChiPSC22) Kit 1 Kit ¥22,051 Human Stem Cell Derived Beta Cells Human Stem Cell Derived Beta Cells Human Stem Cell Derived Beta Cells
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来源于人类iPS细胞的胰腺β细胞
(Pancreatic Beta Cells Derived from Human Induced Pluripotent Stem Cells)
Cellartis® hiPS Beta Cells Kit是由hiPS诱导而来的、通过体外标准化的模拟胚胎发育获得的β细胞。这些细胞分泌大量的胰岛素和C-肽,并且大量表达胰岛素/C-肽、 MAFA、NKX6.1、PDX1、UCN3等蛋白质。完整的试剂盒可以作为理想的实验工具,应用于研究β细胞功能、糖尿病/疾病模型构建、以及胰岛素分泌和调节的化合物筛选。
Cellartis®β细胞是由我们的人iPS细胞系(ChiPSC12;ChiPSC22)分化而来,制备成单细胞悬液,冻存在管中。该试剂盒含有β细胞,包被涂层,基础培养基,以及补充剂。目前提供两种试剂盒供选择,每个试剂盒中的β细胞都来自健康人供体,其中Cellartis hiPS Beta Cells (from ChiPSC22) Kit(Code No. Y10106)的供体细胞是对糖尿病易感的HLA(human leukocyte antigen人类白细胞抗原)类型,HLA-A*02:01。
 
■ 产品特点
· Beta cells from ChiPSC12 and ChiPSC22 originate from healthy human donors
· Beta cells from ChiPSC22 originate from a donor with a diabetes-susceptible HLA type, HLA-A*02:01 (Marron et al. 2002)
· Beta cells from ChiPSC22 can be used alone or in combination with beta cells from ChiPSC12, which have an alternate HLA type
· Cells show protein expression of insulin/C-peptide, MAFA, NKX6-1, PDX1, and UCN3
· Cells demonstrate insulin and C-peptide secretion
· Cells were derived with a protocol that mimics embryonic development
· Cells provide reproducible and biologically relevant data
 
■ 更多信息
Donor information
The tissues used for the reprogramming of somatic cells into hiPSCs are from donors who have signed informed consent which outlines in detail the purpose of the donation and the procedure for processing of the donated tissue. In order to protect the privacy and the confidentiality of the donors, all identifiers associated with the donors have been removed. The donor consent was obtained for commercial use. The donation did not result in any financial gain for the donors.
· ChiPSC12 was sourced from skin fibroblasts from a healthy 24-year-old European/North African male volunteer (77 kg/177 cm).
· ChiPSC22 was sourced from skin fibroblasts from a healthy 32-year-old European/North African male volunteer (74 kg/179 cm) with a diabetes-susceptible HLA type, HLA-A*02:01.
 
HLA typification data for each line is shown below.
Human Stem Cell Derived Beta Cells
 
■ 产品应用
· Disease modeling for diabetes
· Compound screening for insulin secretion and regulation
· Beta-cell functionality
· GSIS (glucose-stimulated insulin secretion) analysis
· Incretin response studies
· Autoimmune beta-cell destruction
· Pancreatitis and beta-cell function
· Transdifferentiation
 
■ 产品组成
· 1 vial (4.8 x 106 viable cells) of Cellartis hiPS Beta Cells (from ChiPSC12) or ChiPSC22
· 11 vials (360 μl/vial) of Cellartis Beta Cell Supplement
· 1 tube (4 ml) of Cellartis Beta Cell Coating
· 1 bottle (160 ml) of Cellartis Beta Cell Basal Medium
· 1 bottle (30 ml) of Cellartis Beta Cell Basal Medium 2
 
 
[Cellartis hiPS Beta Cells (from ChiPSC12) 的胰岛素和 C-肽免疫荧光染色]
Human Stem Cell Derived Beta Cells
解冻后两周的β细胞进行成熟标志胰岛素(A图) 和 C-肽 (B图)免疫荧光染色。合并数据显示了胰岛素和 C-肽共表达细胞 (C图)。细胞核使用DAPI 染色(D图)。
  (图片来源于Takara Bio USA, Inc.)
 
■ 技术资料
Human Stem Cell Derived Beta Cells
Cellartis human iPS cell-derived beta cells for diabetes drug discovery and disease modeling
Studying diabetes and metabolic disorders using hiPSC-derived beta cells.
 
■ 参考文献
Marron, M. et al. Functional evidence for the mediation of diabetogenic T cell responses by HLA-A2.1 MHC class I molecules through transgenic expression in NOD mice. Proc Nat Acad Sci U. S. A. 99, 13753–8 (2002).
 
 
 
■ 产品详情请点击:Human Stem Cell Derived Beta Cells
 
 
 

Human Stem Cell Derived Definitive Endoderm Cells酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Human Stem Cell Derived Definitive Endoderm Cells
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y10040 Cellartis® Definitive Endoderm Cells (from ChiPSC18) 1 Vial ¥8,387 Human Stem Cell Derived Definitive Endoderm Cells Human Stem Cell Derived Definitive Endoderm Cells Human Stem Cell Derived Definitive Endoderm Cells
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Cellartis定型内胚层干细胞
[Cellartis Definitive Endoderm Cells (from ChiPSC18)]
Cellartis定型内胚层干细胞(DE cell)是从hiPS细胞株ChiPSC18经体外诱导分化得来。该产品中,大于90%的细胞显示表达SOX17蛋白。该产品为低温冷冻保存的细胞悬浮形式。本产品适合用于进一步诱导分化成为肝脏细胞类型或是胰脏细胞类型。特别地,Cellartis的肝脏细胞诱导分化试剂盒(Cellartis Hepatocyte Differentiation Kit,Cat. No. Y30050)适合用于DE向肝脏细胞类型的诱导分化工作。
 
■ 产品特点
· Definitive endoderm cells with >90% SOX17-positive cells
· Can be used for generating further-specified cells of endodermal origin, such as hepatocytes or pancreatic endoderm
 
■ 关联产品
Code No. Product Size
Y10052 Cellartis® HEP Coat 3 ml
Y30012 Cellartis® DEF-CS 500 COAT-1 4 ml
T303 iMatrix-511 2 × 175 μg
T304 iMatrix-511 6 × 175 μg
Y30050 Cellartis® Hepatocyte Differentiation Kit 1 Kit
 
■ 产品详情请点击:Human Stem Cell Derived Definitive Endoderm Cells
 
Human Stem Cell Derived Definitive Endoderm Cells
Immunocytochemistry of Cellartis Definitive Endoderm Cells (from ChiPSC18). Cells were thawed, cultured for two days, fixed, and stained for the pluripotency marker OCT4, the definitive endoderm marker SOX17, and the nuclear marker DAPI. Results show that more than 90% of the cell population are definitive endoderm cells and have lost pluripotency.
 
 
 

Human Stem Cell Culture Medium (DEF-CS Xeno-Free)酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Human Stem Cell Culture Medium (DEF-CS Xeno-Free)
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y30045 Cellartis® DEF-CS 500 Xeno-Free Culture Medium w/o antibiotics 1 Kit ¥2,923 Human Stem Cell Culture Medium (DEF-CS Xeno-Free) Human Stem Cell Culture Medium (DEF-CS Xeno-Free) Human Stem Cell Culture Medium (DEF-CS Xeno-Free)
Cellartis Y30047 Cellartis® DEF-CS 500 Xeno-Free 3D Spheroid Culture Medium 1 Kit ¥3,747 Human Stem Cell Culture Medium (DEF-CS Xeno-Free) Human Stem Cell Culture Medium (DEF-CS Xeno-Free) Human Stem Cell Culture Medium (DEF-CS Xeno-Free)
Cellartis Y30071 Cellartis® DEF-CS 500 Xeno-Free GMP Grade Basal Medium (Prototype) 500 ml ¥6,088 Human Stem Cell Culture Medium (DEF-CS Xeno-Free) Human Stem Cell Culture Medium (DEF-CS Xeno-Free) Human Stem Cell Culture Medium (DEF-CS Xeno-Free)
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临床前扩增人干细胞培养基系列(Xeno-free)
(The Xeno-Free Culture Media Product Family for Preclinical Expansion of Human iPS Cells)
Cellartis DEF-CS (Xeno-free) 培养基系列,完全无任何人源及动物源成分,被用于人类诱导多能干细胞 (iPSCs) 扩增。该款产品不使用饲养层 (feeder),而是使用xeno-free的基质物,比如 新型的iMatrix-511。使用DEF-CS 培养基 (xeno-free) 培养的细胞,拥有高的增殖率和稳定的核型,允许2D-单层培养和3D-悬浮培养进行有效扩增。
 
这些产品主要适用于
· 基础研究和临床前概念研究—Cellartis DEF-CS 500 Xeno-Free Culture Medium帮助您从基础研究过渡到下一阶段相关疾病模型的验证和生产制造战略的开展。
 
· 扩增到足够的细胞数—Cellartis DEF-CS 500 Xeno-Free 3D Spheroid Culture Medium搭配灌注生物反应器系统,提供了一个产生大量高质量的、未分化的、用作起始细胞治疗材料的理想方法。
 
· GMP 级生产和下游使用于临床研究—Cellartis DEF-CS 500 Xeno-Free GMP Grade Basal Medium (Prototype)遵循严格的生产准则,确保没有可能损害患者的人类和动物来源的物质。
 
■ 产品特点
Cellartis DEF-CS 500 Xeno-Free Culture Medium
· Maintains pluripotency and stable karyotype in a 2D non-colony monolayer culture
· Ideal for maintenance of undifferentiated pluripotent cells and scale-up to therapeutically relevant cell numbers
· Fully compatible with the GMP-grade version for a seamless transition to the clinic
Cellartis DEF-CS 500 Xeno-Free 3D Spheroid Culture Medium
· Pluripotent cells in the spheroids retain full differentiation potential and stable karyotype
· Produces 5 × 109 cells in a 1 L bioreactor within 3–4 passages
· Provides a distinct advantage for scale-up to therapeutically relevant cell numbers
Cellartis DEF-CS 500 Xeno-Free GMP Grade Basal Medium (Prototype)
· Optimized for small- or large-scale production of pluripotent cells for downstream use in clinical research settings
· Manufactured as a quality-assured product, according to the guidelines for GMP for investigational products
· Rigorous manufacturing standards ensure quality and consistency; all materials are of clinical quality, with traceable production processes
 
■ 产品应用
· Culturing human iPS cells in completely xeno-free conditions for preclinical studies.
 
■ 产品目录
Code No. Product Size
Y30045 Cellartis® DEF-CS 500 Xeno-Free Culture Medium w/o antibiotics 1 kit
Y30042 Cellartis® DEF-CS 500 Xeno-Free Additives 1 kit
Y30046 Cellartis® DEF-CS 500 Xeno-Free Basal Medium w/o antibiotics 500 ml
 
Code No. Product Size
Y30047 Cellartis® DEF-CS500 Xeno-Free 3D Spheroid Culture Medium w/o antibiotics 1 kit
Y30046 Cellartis® DEF-CS 500 Xeno-Free Basal Medium w/o antibiotics 500 ml
Y30048 Cellartis® DEF-CS500 Xeno-Free 3D Spheroid Additives 1 kit
 
Code No. Product Size
Y30071 Cellartis® DEF-CS 500 Xeno-Free GMP Grade Basal Medium (Prototype) 500 ml
 
■ 关联产品
Code No. Product Size
T303 iMatrix-511 2 × 175 μg
 
■ 产品详情请点击
Y30045:Human Stem Cell Culture Medium (DEF-CS Xeno-Free)
Y30047:Human Stem Cell Culture Medium (DEF-CS Xeno-Free)
Y30071:Human Stem Cell Culture Medium (DEF-CS Xeno-Free)
 
Human Stem Cell Culture Medium (DEF-CS Xeno-Free)
High-quality hiPS cell aggregates grown with Cellartis DEF-CS 500 Xeno-Free 3D Spheroid Culture Medium in stirred tank bioreactors (in perfusion mode). hiPS cell aggregates grown under 4% O2 perfusion conditions were sequentially passaged three times by mechanical disruption (protocol adapted from Otsuji et al. 2014); cells were counted at passage on days 4 and 7, and at the final time point on day 11, to determine the expansion factor relative to the initial seeding number.
 
Human Stem Cell Culture Medium (DEF-CS Xeno-Free)
High-quality hiPS cell aggregates were grown with Cellartis DEF-CS 500 Xeno-Free 3D Spheroid Culture Medium in stirred tank bioreactors (in perfusion mode). After expansion of hiPS cells as aggregates, cells were harvested on day 4 and expression of OCT4, SSEA-4, TRA-1-60 (pluripotency markers) and SSEA-1 and SOX17 (differentiation markers) was analyzed by immunostaining.
 
  Takara Bio USA, Inc.比较结果

Mouse ES and iPS cell culture medium (2i)酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Mouse ES and iPS cell culture medium (2i)
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y40031 Cellartis® 2i mES/iPSC Culture Medium 200 ml ¥3,158 Mouse ES and iPS cell culture medium (2i) Mouse ES and iPS cell culture medium (2i) Mouse ES and iPS cell culture medium (2i)
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小鼠ES/iPS细胞培养基(2i Medium)
Cellartis 2i mES/iPSC Culture Medium是一款无血清的、成分确定的细胞培养基,可用于小鼠ES和iPS细胞建系。该培养基含有两种小分子抑制剂(基于2i技术:CHIR99021、PD0325901),分别是用于抑制GSK3β和ERK/MEK介导的分化诱导信号通路,以及促进细胞增殖。
单独使用Cellartis 2i mES/iPSC Culture Medium,可以长期维持小鼠ES细胞处于原始状态和基态(Naive, ground state)或者小鼠iPS细胞处于完全多能性(Fully pluripotent)的状态。添加LIF(Leukemia Inhibitory Factor),可以通过调节Nanog信号通路使得Partial-iPS细胞或Pre-iPS细胞有效转化为具有完全多能性的iPS细胞(Silva et al. 2008; Silva et al. 2009).
2i技术也应用于从难获得具有生殖系能力(germline-competent)ES细胞的小鼠品系中建立ES细胞系(Nichols et al. 2009),以及用于从人类和家畜物种中诱导iPS细胞(Wang et al. 2011; Malaver-Ortega et al. 2012)。
 
■ 产品特点
· 成分确定、无血清,促进细胞增殖并维持多能性状态
· 通过添加小分子抑制剂来阻断GSK3β和 ERK/MEK介导的分化诱导通路,并维持小鼠ES细胞和iPS细胞长期具备生殖系能力
· 添加 LIF,使Partial-iPS细胞或Pre-iPS细胞有效转化为具有完全多能性的iPS细胞
 
■ 产品应用
· 建立小鼠ES细胞系和iPS细胞系
· 长期维持纯的ES细胞或iPS细胞
· 使Partial-iPS细胞或Pre-iPS细胞转化为具有完全多能性的iPS细胞(需添加 LIF)
 
■ 产品组成
· 1 bottle (200 ml) of Cellartis mES/iPSC Culture Basal Medium
· 1 tube (200 μl) Cellartis 2i mES/iPSC Supplement
 
 
■ 产品详情请点击:Mouse ES and iPS cell culture medium (2i)
 
■ 参考文献
1. Malaver-Ortega, L. F. et al. The state of the art for pluripotent stem cells derivation in domestic ungulates. Theriogenology 78, 1749–1762 (2012).
2. Nichols, J. et al. Validated germline-competent embryonic stem cell lines from non-obese diabetic mice. Nat. Med15, 814–818 (2009).
3. Silva, J. et al. Promotion of reprogramming to ground state pluripotency by signal inhibition. PLoS Biol. 6, e253 (2008).
4. Silva, J. et al. Nanog is the gateway to the pluripotent ground state. Cell 138, 722–37 (2009).
5. Wang, W. et al. Rapid and efficient reprogramming of somatic cells to induced pluripotent stem cells by retinoic acid receptor gamma and liver receptor homolog. PNAS 108, 18283–18288 (2011).
 

Mouse ES and iPS cell culture medium (3i)酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Mouse ES and iPS cell culture medium (3i)
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y40011 Cellartis® 3i mES/iPSC Culture Medium 200 ml ¥4,226 Mouse ES and iPS cell culture medium (3i) Mouse ES and iPS cell culture medium (3i) Mouse ES and iPS cell culture medium (3i)
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小鼠ES/iPS细胞培养基(3i Medium)
Cellartis 3i mES/iPSC Culture Medium是一款无血清的、成分确定的细胞培养基,可用于小鼠ES细胞的建系、多能性维持和增殖培养。该培养基含有三种小分子抑制剂(基于3i技术:ERK/MEK inhibitor、GSK3β inhibitor、FGFR-TK inhibitor),用于抑制分化诱导信号通路和促进细胞增殖,以及可以在无需额外添加细胞因子刺激的情况下维持多能性基态(Pluripotent ground state)。因此,Cellartis 3i mES/iPSC Culture Medium可以应用于多能性分子机制研究和分析。
Cellartis 3i mES/iPSC Culture Medium也应用于从标准条件下难建立ES细胞的小鼠品系中建立ES细胞系(Ying et al. 2008)。相较于LIF(Leukemia Inhibitory Factor)+血清或BMP(Bone Morphogenetic Protein)条件下培养的小鼠ES细胞,使用Cellartis 3i mES/iPSC Culture Medium培养的小鼠ES细胞可以持续地以双倍速度扩增 (Ying et al. 2008; Kiyonari et al. 2010; Iijima et al. 2010)。此外,同样与LIF+血清或BMP条件下培养的小鼠ES细胞相比,使用Cellartis 3i mES/iPSC Culture Medium培养的小鼠ES细胞表现出更高的多能性克隆形成效率(Clonogenicity)。
 
■ 产品特点
· 成分确定、无血清、无饲养层、无细胞因子
· 通过添加小分子抑制剂来抑制分化诱导通路并维持小鼠ES细胞处于多能性状态
· 支持小鼠ES细胞增殖并维持≥99%纯度
 
■ 产品应用
· 维持小鼠ES细胞处于多能性状态
· 从难建系的小鼠品系中制备小鼠ES细胞
· 高效制备具有完全多能性的小鼠ES细胞
 
■ 产品组成
· 1 bottle (200 ml) of Cellartis mES/iPSC Culture Basal Medium
· 1 tube (200 μl) of Cellartis 3i mES/iPSC Supplement
 
 
■ 产品详情请点击:Mouse ES and iPS cell culture medium (3i)
 
Cellartis 3i mES/iPSC Culture Medium维持小鼠ES细胞处于同质化、未分化状态。
Mouse ES and iPS cell culture medium (3i)
小鼠ES细胞系ES-E14TG2a分别使用Cellartis 3i mES/iPSC Culture Medium(A)和 GMEM-based medium(B)培养,通过碱性磷酸酶(Alkaline phosphatase, AKP)染色检测ES细胞多能性,左边为染色前,右边为AKP染色后结果,可以看出Cellartis 3i Medium培养的小鼠ES细胞为均一的、未分化细胞。
 
Cellartis 3i mES/iPSC Culture Medium培养建立的小鼠ES细胞系具备高嵌合率。
Mouse ES and iPS cell culture medium (3i)
将ES细胞注射并植入胚胎后,通过出生小鼠个体的毛色来判断ES细胞的嵌合率。从Cellartis 3i medium培养建立的ES细胞所获得的小鼠均为黑色毛,即在单代内就获得了完全是ES细胞来源的嵌合体小鼠(A),具备高嵌合率。而从常规培养基培养建立的ES细胞所获得的嵌合体小鼠毛色混杂,未完全嵌合(B)。
 
■ 参考文献
1. Ying, Q. L. et al. The ground state of embryonic stem cell self-renewal. Nature 453, 519–23 (2008).
2. Kiyonari, H. et al. Three inhibitors of FGF receptor, ERK, and GSK3 establishes germline-competent embryonic stem cells of C57BL/6N mouse strain with high efficiency and stability. Genesis 48, 317–327 (2010).
3. Iijima, S. et al. Effect of different culture conditions on establishment of embryonic stem cells from BALB/cAJ and NZB/BINJ mice. Cell Reprogram. 12, 679–688 (2010).
 
 

Adherent Neural Stem Cell Culture Media酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Adherent Neural Stem Cell Culture Media
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y40000 RHB-Basal® 500 ml ¥2,259 Adherent Neural Stem Cell Culture Media Adherent Neural Stem Cell Culture Media Adherent Neural Stem Cell Culture Media
Cellartis Y40001 RHB-A® 500 ml ¥3,627 Adherent Neural Stem Cell Culture Media Adherent Neural Stem Cell Culture Media Adherent Neural Stem Cell Culture Media
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无标题文档

Neural Stem Cell Culture Media
RHB-A和RHB-Basal是两款无血清、成分完全确定的培养基,用于人类或小鼠神经干细胞(NS)的定向分化获取、维持培养、扩增、诱导分化成神经细胞等用途。RHB-A可用来培养扩增贴壁的人或小鼠的神经干细胞。在RHB-A中培养的神经干细胞保持着稳定的神经干细胞分化发育成为成熟的神经细胞的能力 (1-3) 。另外,RHB-A还用来定向分化小鼠胚胎干细胞(mouse ES)至神经前体细胞 (neural precursors) (4-6)。
RHB‐Basal不包含生长因子或是神经细胞相关的添加物。因此,可以RHB‐Basal为基础,通过添加客户认为需要的添加物来开发适合客户的特殊类型细胞的培养基。
 
■ 产品特点
· RHB-A is a proprietary, fully defined, and serum-free medium designed to maintain pure populations of adherent human and mouse NS cells
· RHB-Basal medium is animal component-free and contains no neuronal supplements; the media can be customized by the addition of supplements
 
■ 产品应用
· Derivation of mouse and human NS cells from ES cells and fetal and adult tissues
· Maintenance and propagation of adherent mouse and human NS cells
· Differentiation of mouse and human NS cells into functional neurons
· Differentiation of mouse ES cells to neuronal precursors
· Refer to the Data Sheet for additional examples of use
 
■ 产品详情请点击:Adherent Neural Stem Cell Culture Media
 
Adherent Neural Stem Cell Culture Media
Adherent mouse neural stem cells cultured in RHB-A medium supplemented with epidermal growth factor and fibroblast growth factor-2 express neural stem cell markers, including Nestin, Vimentin (3CB2), radial glial cell marker-2 (RC2), glial fibrillary acidic protein (GFAP), and microtubule associate protein (MAP).
 
参考文献:
1. Ying QL, et al. (2003) Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture. Nature Biotechnology 21:183-186.
2. Conti L, et al. (2005) Niche-Independent symmetrical self-renewal of a mammalian tissue stem cell. PLoS Biology 3(9):e283.
3. Pollard SM, et al. (2006) Adherent Neural Stem (NS) cells from fetal and adult forebrain. Cerebral Cortex 16:112-120. 4. Diogo MM, et al. (2008) Optimization and integration of expansion and neural commitment of mouse embryonic stem cells. Biotechnology and Applied Biochemistry 49:105-112.
5. Pollard SM, et al. (2008) Fibroblast growth factor induces a neural stem cell phenotype in foetal forebrain progenitors and during embryonic stem cell differentiation. Molecular and Cellular Neuroscience 38:393:403.
6. Sun Y, et al. (2008) Long-term tripotent differentiation capacity of human neural stem (NS)cells in adherent culture. Molecular and Cellular Neuroscience 38:245-258.
7. Pollard SM, et al. (2009) Glioma stem cell lines expanded in adherent culture have tumor-specific phenotypes and are suitable for chemical and genetic screens. Cell Stem Cell 4:568-580.
8. Abranches E, et al. (2009) Neural differentiation of embryonic stem cells in vitro: A road map to neurogenesis in the embryo. PLoS ONE 4(7): e6286.
9. Fernandes et al. (2010) Hypoxia enhances proliferation of mouse embryonic stem cell-derived neural stem cells. Biotechnology and Bioengineering 106: 260–270.
10. Fernandes, T.G., et al. (2010) Different stages of pluripotency determine distinct patterns of proliferation, metabolism, and lineage commitment of embryonic stem cells under hypoxia. Stem Cell Research 5(1):76-89.
11. Fernandes et al. (2010) Three-dimensional cell culture microarray for high-throughput studies of stem cell fate. Biotechnology and Bioengineering 106: 106-118.
12. Hansen, et al. (2010) Neurogenic radial glia in the outer subventricular zone of human neocortex. Nature doi: 10.1038/nature08845.
13. Hook L, et al. (2011) Non-immortalized human neural stem (NS) cells as a scalable platform for cellular assays. Neurochem Int. 59(3): 432-44.
 
 

Mouse Feeder Cell Quantification Kit酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Mouse Feeder Cell Quantification Kit
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Takara RR290 Mouse Feeder Cell Quantification Kit 100 Rxns Inquire Mouse Feeder Cell Quantification Kit Mouse Feeder Cell Quantification Kit Mouse Feeder Cell Quantification Kit
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无标题文档

 
 
■ 制品内容(25 μl × 100次量)
1. TB Green Premix Ex Taq II(Tli RNaseH Plus)(2×Conc.) 625 μl × 4
2. Mouse_Mt_primer(10 μM each) 100 μl
3. Mouse_genome_primer(10 μM each) 100 μl
4. Human_primer_Type I(CHR3)(10 μM each) 100 μl
5. Human_primer_Type II(CHR7)(10 μM each) 100 μl
6. Human_primer_Type III(CHR15)(10 μM each) 100 μl
7. dH2O 1 ml×2
8. EASY Dilution(for Real Time PCR) 1 ml×2
9. ROX Reference Dye(50×Conc.) 50 μl
10. ROX Reference Dye II(50×Conc.) 50 μl
11. Human Genomic DNA(100 ng/μl) 20 μl
 
*:用于Applied Biosystems的Real Time PCR仪,用以校正孔与孔之间的荧光信号误差。
◆ 使用ROX Reference Dye 的PCR仪
·StepOnePlus Real-Time PCR System(Thermo Fisher Scientific)
◆ 使用ROX Reference Dye II的PCR仪
·Applied Biosystems 7500 Fast Real-Time PCR System(Thermo Fisher Scientific)
◆ 不使用ROX Reference Dye或ROX Reference Dye II的PCR仪
·Thermal Cycler Dice Real Time System II/Lite (Code No. TP900/TP960/TP700/TP760: 终卖)
 
Limited Use Label License:[M82]
 
■ 制品说明
小鼠饲养细胞可用于维持包括hESC(人胚胎干细胞)及 hiPSC(人诱导多能干细胞)等干细胞的增殖及分化。但对于干细胞的某一种应用如再生医学,去除异种饲养细胞是非常关键的。为了保证干细胞质量,有必要建立残留小鼠细胞的检测体系。
本制品以从hESC及hiPSC等培养干细胞提取的基因组DNA为模板,应用Real Time PCR方法对残留的小鼠饲养细胞来源的基因组DNA进行高灵敏度检测和定量的试剂盒。
本制品使用的引物是在小鼠线粒体(mt)DNA区域设计的Real Time PCR扩增用引物,可高灵敏度地检测小鼠饲养细胞来源的基因组DNA。现已知mtDNA拷贝数依据细胞种类、细胞株及分化状态不同而不同,使用的小鼠饲养细胞批次间也可能有差异,因此在试剂盒中附带了可检测小鼠mtDNA拷贝数的引物,用于小鼠饲养细胞批次间的校正。
*:使用同一个标准曲线对多批次小鼠饲养细胞来源的基因组DNA定量时,需确认各批次间的mtDNA拷贝数无差异。当小鼠饲养细胞每个批次都制作标准曲线时,则不需要校正。
 
■ 保 存
TB Green Premix Ex Taq II(Tli RNaseH Plus)

4℃可保存6个月。
避光保存。避免污染。
※长期保存时,请于-20℃保存。产品融解后请于4℃保存,并在6个月内用完。

其他组分

-20℃保存
 
 

Neural Progenitor Cell Antibodies酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Neural Progenitor Cell Antibodies
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Takara M194 Anti-Mouse L7/Pcp2, Polyclonal 0.2 mg ¥3,244 Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies
Takara M195 Anti-Mouse Otp, Polyclonal 0.2 mg ¥3,244 Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies
Takara M196 Anti-Mouse Emx1, Polyclonal 0.2 mg ¥3,244 Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies
Takara M198 Anti-Mouse Otx2, Polyclonal 0.2 mg ¥3,244 Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies
Takara M202 Anti-Human L7/Pcp2, Polyclonal 0.2 mg ¥3,244 Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies
Takara M227 Anti-Human/Mouse Bf1, Polyclonal 0.2 mg ¥3,244 Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies
Takara M228 Anti-Mouse Rx, Polyclonal 0.2 mg ¥3,244 Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies
Takara M229 Anti-Human/Mouse Rx, Polyclonal (Guinea Pig) 0.2 mg ¥3,244 Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies
Takara M230 Anti-Mouse Irx3, Polyclonal 0.2 mg ¥3,275 Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies
Takara M231 Anti-Human Crx, Polyclonal 0.2 mg ¥3,244 Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies Neural Progenitor Cell Antibodies
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无标题文档

Antibodies to Neural Progenitor Cells: Bf1, Crx, Emx1, Irx3,
L7/Pcp2, Otp, Otx2 and Rx

神经前体细胞(Neural progenitor cells)表达特异性的蛋白标志物,因此,这些标志物的抗体被应用于干细胞分化过程的研究,比如胚胎发育和器官发育。在再生医学研究中,这些抗体被用做干细胞神经分化的标记物。在免疫组织化学中,这些抗体用于组织辨别和Western blotting。
 
Antibody to Cell type detected Antigen Species Specificity Code No.
Bf1 Cerebral neural progenitor cells in the telencephalon C-terminal peptide of human/mouse Bf1 conjugated with bovine thyroglobulin Human
Mouse
C-terminal region of human and mouse Bf1 M227
Crx Retinal photoreceptor cells(cone and rod cells) during embryonic development KLH-conjugated peptide fragments of human Crx Human Human Crx M231
Emx1 Cerebral cortex neurons during embryonic development KLH-conjugated peptide of mouse Emx1 Mouse C-terminal region of mouse Emx1 M196
Irx3 Neural plate progenitor cells involved in caudal nerve development KLH-conjugated peptide from the C-terminal region of mouse Irx3 Mouse Mouse Irx3 M230
L7/Pcp2 Purkinje progenitor cells KLH-conjugated peptide from the C-terminal region of L7/Pcp2 Mouse C-terminal region of mouse L7/Pcp2 M194
Human C-terminal region of mouse L7/Pcp2; Reacts with human & mouse L7/Pcp2 M202
Otp Hindbrain and hypothalamic neurons during embryonic development KLH-conjugated peptide from the C-terminal region of mouse Otp Mouse C-terminal region of mouse Otp M195
Otx2 Retinal photoreceptor cells during embryonic development KLH-conjugated peptide from the C-terminal region of Otx2 Mouse C-terminal region of mouse Otx2 M198
Rx Retinal progenitor cells KLH-conjugated peptide fragments of Rx Mouse Mouse Rx M228
Human
Mouse
Human & mouse Rx M229
 
■ 产品应用
· Immunohistochemical staining on fixed tissue
· Western blotting under reducing conditions (Cat. # M196, M228, M229, M230, M231)
 
Neural Progenitor Cell Antibodies
Immunohistochemical Staining with Polyclonal Antibody to Mouse Otx2 (Cat. No. M198) and DAPI. Tissue: Embryonic mouse, day 10 (E10), forebrain to midbrain. Fixation: 4% paraformaldehyde, 6 h. Permeabilization: 0.3% Triton X-100 in PBS. Blocking: 2% skim milk in PBS. Blue, DAPI stain; red, anti-Otx2 antibody.
 

Pluripotent Cell Detection酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Pluripotent Cell Detection
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y20010 hES-Cellect 150 μg ¥6,395 Pluripotent Cell Detection Pluripotent Cell Detection Pluripotent Cell Detection
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检测多能性细胞用抗体
(Monoclonal Antibodies for Detecting Pluripotent Cells)
Cellect系列单克隆抗体用于鉴别、分析、分离具有发育多能性的人类胚胎干细胞(ES)和诱导干细胞(iPS)。本系列抗体特异性地识别具有发育多能性的干细胞表面的抗原,而这些抗原在干细胞定向分化后会被弱化调控。
 
■ 产品特点
· hES-Cellect is high-purity monoclonal antibodies that recognize surface markers specific to pluripotent cells
· hES-Cellect can be used without cell fixation or permeabilization to detect and select live cells
 
Product Name Human ES Cells Human iPS Cells
hES-Cellect × ×
 
■ 产品应用
· Identification and/or isolation of human ES and iPS cells by immunocytochemistry, flow cytometry,and magnetic cell separation
· Separation of human pluripotent stem cells from feeder cells and differentiated progeny
· Early identification of reprogrammed human iPS cell clones
 
 
■ 产品详情请点击:Pluripotent Cell Detection
 

Human Stem Cell Markers酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Human Stem Cell Markers
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Takara M221 Anti-Human Oct4, Monoclonal 0.1 mg Inquire Human Stem Cell Markers Human Stem Cell Markers Human Stem Cell Markers
Takara M222 Anti-Human Lin28, Monoclonal 0.1 mg Inquire Human Stem Cell Markers Human Stem Cell Markers Human Stem Cell Markers
Takara M223 Anti-Human Sox2, Monoclonal 0.1 mg Inquire Human Stem Cell Markers Human Stem Cell Markers Human Stem Cell Markers
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人干细胞标志物抗体
(Human Stem Cell Markers)
通过向体细胞引入“发育全能性相关因子”可以获得诱导干细胞(iPS)。例如,通过在人类体细胞内异位表达(ectopic expression)Oct4, Sox2, Lin28以及 Nanog,可以诱导细胞获取发育全能性(pluripotency)(Yu et al., 2007)。针对这些因子的抗体,可以使用在Western blot实验或是免疫组织化学实验中。
 
■ 产品应用
· Western blot analysis under reducing conditions
· Immunocytochemistry
 
Human Stem Cell Markers
 
参考文献:
Yu, J. et al. (2007) Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells. Science 318:1917–1920.
 

iPS Cell to Endothelial Cell Differentiation System酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

iPS Cell to Endothelial Cell Differentiation System
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y50300 MiraCell® iPS Cell to Endothelial Cell Differentiation Kit 1 Kit ¥13,273 iPS Cell to Endothelial Cell Differentiation System iPS Cell to Endothelial Cell Differentiation System iPS Cell to Endothelial Cell Differentiation System
Cellartis Y50301 MiraCell® iPS Cell to Endothelial Cell Differentiation Kit with DEF-CS 500 Culture System 1 Kit ¥18,627 iPS Cell to Endothelial Cell Differentiation System iPS Cell to Endothelial Cell Differentiation System iPS Cell to Endothelial Cell Differentiation System
Cellartis Y50053 MiraCell® EC Culture Medium 500 ml ¥4,148 iPS Cell to Endothelial Cell Differentiation System iPS Cell to Endothelial Cell Differentiation System iPS Cell to Endothelial Cell Differentiation System
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iPS向血管内皮细胞定向分化系统
(iPS Cell to Endothelial Cell Differentiation System)
MiraCell iPS Cell to Endothelial Cell Differentiation Kit用于人iPS细胞向血管内皮细胞定向分化诱导,试剂盒中包含2种包被剂、2种分化用基础培养基和6种培养基添加剂,足够支持1个T25培养瓶 (1.5×105 Cells)或3个6孔板单孔(1.8×105 cells)的iPS细胞进行内皮方向分化诱导,分化周期为12天(Day0至Day11)。建议分化起始的iPS细胞先使用Cellartis® DEF-CS 500 Culture System(Y30010)驯化3-5代,以确保iPS细胞处于良好的多能性状态。
诱导分化获得的高纯度(CD31阳性率>80%*)血管内皮细胞,经鉴定表达CD31、CD144、Tie2、VEGFR2等血管内皮标志物和具备小管(Tube)形成能力。分化好的血管内皮细胞(Day12以后)可以使用MiraCell EC Culture Medium(Y50053)长期扩大培养,也可以冷冻保存备用。(*起始的人iPS细胞株不同,得到的血管内皮细胞性能可能不同。)
 
本产品使用京都大学iPS细胞研究所开发的人血管内皮细胞的制备技术,由Takara和iHeart Japan公司共同研发,由Takara制造并销售。
 
■ 产品特点
· 完整的试剂盒,支持人iPS细胞-中胚层细胞-血管内皮细胞定向分化
· 多个iPS细胞株测试验证,支持高效分化诱导(>80%)
· 无需基因修饰和药物筛选过程,操作简便
· 获得的血管内皮细胞,可冷冻保存和扩大培养
 
■ 产品组成
MiraCell® iPS Cell to Endothelial Cell Differentiation Kit,Y50300
· Coating Reagent 1 (Day0 & 2);Coating Reagent 2 (Day11)
· EC Differentiation Basal Medium 1 (Day3~9);EC Differentiation Basal Medium 2 (Day11)
· Supplement 1 (Day3);Supplement 2 (Day4);Supplement 3 (Day7);Supplement 4 (Day8);Supplement 5 (Day9);Supplement 6 (Day11)
 
iPS Cell to Endothelial Cell Differentiation System
 
MiraCell® iPS Cell to Endothelial Cell Differentiation Kit with DEF-CS 500 Culture System,Y50301
· MiraCell® iPS Cell to Endothelial Cell Differentiation Kit,Y50300
· Cellartis® DEF-CS 500 Culture System,Y30010
 
iPS Cell to Endothelial Cell Differentiation System
 
■ 实验例
iPS Cell to Endothelial Cell Differentiation System
图1. 3种人iPS细胞株使用MiraCell iPS Cell to Endothelial Cell Differentiation Kit分化诱导,获得的血管内皮细胞的形态比较(A)和标志蛋白质(B)比较。
 
iPS Cell to Endothelial Cell Differentiation System
图2. 3种人iPS细胞株使用MiraCell iPS Cell to Endothelial Cell Differentiation Kit分化诱导,获得的血管内皮细胞继续使用MiraCell EC Culture Medium维持培养,可长时间维持高纯度。()内的数值表示Day32的细胞扩大倍数,以Day8的细胞数为1。
 
iPS Cell to Endothelial Cell Differentiation System
图3. 冷冻保存的、使用MiraCell iPS Cell to Endothelial Cell Differentiation Kit分化诱导的血管内皮细胞(Day16),在解冻复苏培养Day8后的小管形成实验结果。
 
 
 
产品详情请点击:http://catalog.takara-bio.co.jp/product/basic_info.php?unitid=U100009383
 
 

Human iPS Cell Editing Systems酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Human iPS Cell Editing Systems
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Cellartis Y30021 Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit 1 kit ¥7,860 Human iPS Cell Editing Systems Human iPS Cell Editing Systems Human iPS Cell Editing Systems
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无标题文档

 
从开始到结束:人iPS细胞基因编辑和单细胞克隆系统
Start-To-Finish hiPSC Editing and Single-Cell-Cloning Systems
人诱导多能干细胞(Human induced pluripotent stem cells,hiPSCs)是一个功能强大的研究工具和疾病模型,因为其具备增殖、自我更新、分化为多种细胞类型的能力。此外,人iPS细胞还可以再现细胞来源个体的疾病表型和基因型。将人iPS细胞结合基于CRISPR/Cas9的基因编辑技术,可以在同基因的细胞系中研究特定基因改变引起的功能影响,因为基因编辑过的人iPS细胞能提供可再生的、无遗传变异的患病细胞和健康细胞资源。
CRISPR/Cas9系统已经成为一个强大的基因编辑工具,因为其高的靶向特异性,编辑效率和易用性。该技术的强大主要源于它的简单,因为全部只需要一个Cas9核酸酶结合一个单链向导RNA(single guide RNA,sgRNA) 就可以决定靶向特异性(Jinek et al. 2012)。这种RNA编程的(RNA-programmable)方法利用了非同源末端连接(non-homologous end joining,NHEJ) DNA修复途径的易错特性,进而产生基因敲除(通过插入/删除)。这种方法也可以通过同源重组(homology-directed repair,HDR)途径被用于基因敲入。
 
CRISPR/Cas9系统的组件可以通过多种方法成功导入至靶细胞,包括基于载体的表达系统,RNA转染, 导入Cas9/sgRNA核糖核蛋白(ribonucleoprotein,RNP)复合物(Sander and Joung 2014)等。与基于载体的方法相比,直接导入Cas9/sgRNA RNPs提供了一种快速转入基因编辑实验的方法,同时脱靶效应的可能性更小(Kim et al. 2014)。
 
一旦Cas9/sgRNA RNP复合物被导入,为了分离和筛选感兴趣的基因型,单细胞必须被分离并扩增为克隆细胞系。传统上,从基因编辑的hiPS细胞建立一个克隆群是低效的、具有挑战性的和耗时的,因为hiPS 细胞是以集落生长和传代的。通常,这会导致细胞死亡和提前分化。然而,Cellartis单细胞克隆系统包含一个成分确定的培养系统(由基础培养基,包被剂,和添加剂组成),用于有效形成单细胞克隆和扩增基因编辑后的hiPSC克隆。DEF-CS culture system (Asplund et al. 2016)是一个基于单层培养的系统,规避了集落状培养带来的挑战,允许单细胞传代和促进接种的单细胞存活和后续扩增。
 
Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit (Code No. Y30021) 是一个完整的,在无饲养层和成分确定条件下,用于hiPSCs有效扩增和扩大生产的系统。试剂盒包含了从接种单细胞至96孔板到扩增至48孔板过程中所有必要的试剂。单细胞克隆扩增后,hiPSCs可以使用Cellartis DEF-CS 500 Culture System (Code No. Y30010)继续培养。
 
关联基因编辑系统:
一、电穿孔法基因编辑
通过电穿孔的方法对hiPSCs进行有效的基因编辑,Takara提供重组Cas9蛋白质和产生大量sgRNAs所有必要的试剂,用于电穿孔hiPSCs。后续与Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit (Code No. Y30021)一起使用,用于进行基因编辑后的单细胞克隆形成和扩增。单细胞克隆扩增后,hiPSCs可以使用Cellartis DEF-CS 500 Culture System (Code No. Y30010)继续培养。
二、Gesicle法基因编辑
Gesicles法是一种无毒的、高效的、替代电穿孔的方法,不依赖昂贵的设备或耗材。通过Gesicle法导入Cas9/sgRNA复合物进行有效的基因编辑,Takara提供所有的必要试剂用于产生细胞衍生的纳米囊泡(Gesicles),来转导Cas9和sgRNA至hiPSCs。后续可与Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit (Code No. Y30021)一起使用,用于进行基因编辑后的单细胞克隆形成和扩增。单细胞克隆扩增后,hiPSCs可以使用Cellartis DEF-CS 500 Culture System (Code No. Y30010)继续培养。
 
■ 产品特点
· Highly efficient gene editing in hiPSCs—use either electroporation or gesicles to deliver Cas9 protein and sgRNA with no genomic integration and reduced off-target effects
· Superior single-cell survival—edited hiPSCs exhibit high (typically ~50%) survival when seeded as single cells in a 96-well plate
· Maintenance of pluripotency after editing and during single-cell cloning—hiPSCs maintain high levels (>90%) of pluripotency markers Oct-4, TRA-1-60, and SSEA-4
· Stable karyotype from start to finish—maintain a normal and stable karyotype throughout editing, single-cell cloning, and expansion
· Flexibility to perform gene editing experiments your way—choose from complete kits that provide editing and single-cell cloning solutions using either electroporation-based or utilize the culture media kit (Code No. Y30021) to perform efficient single-cell cloning following your own editing experiments
· Continuous use of DEF-CS technology throughout gene editing/single-cell cloning experiments—use the Cellartis DEF-CS 500 Culture System (Code No. Y30010) before and during gene editing experiments, then during scale-up after single-cell cloning experiments
 
■ 产品组成
Cellartis® iPSC Single-Cell Cloning DEF-CS Culture Media Kit
(Code No. Y30021,1 kit)
· Cellartis DEF-CS 500 Basal Medium (Code No. Y30011; 500 ml)
· Cellartis iPSC Single-Cell Cloning DEF-CS COAT-1 (Code No. Y30018; 2 × 800 μl)
· Cellartis iPSC Single-Cell Cloning DEF-CS Additives
   (Code No. Y30019; 2 × 750 μl, 1 × 500 μl, 1 × 500 μl)
 
关联基因编辑产品
电穿孔法基因编辑
Guide-it™ sgRNA In Vitro Transcription Kit (632635; 1 kit)
Guide-it™ Recombinant Cas9 (Electroporation-Ready) (632641; 100 μg)
Gesicle法基因编辑
Guide-it™ CRISPR/Cas9 Gesicle Production System (632613;1 kit)
 
■ 产品应用
· CRISPR/Cas9-mediated gene editing of hiPSCs
· Single-cell cloning of hiPSCs
· Disease modeling
 
■ 参考文献
Asplund, A. et al. One Standardized Differentiation Procedure Robustly Generates Homogenous Hepatocyte Cultures Displaying Metabolic Diversity from a Large Panel of Human Pluripotent Stem Cells. Stem Cell Rev. Reports 12, 90-104 (2016).
Jinek, M. et al. A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science 337, (2012).
Kim, S., Kim, D., Cho, S. W., Kim, J. & Kim, J.-S. Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins. Genome Res. 24, 1012-9 (2014).
Sander, J.-D. & Joung, J.-K. CRISPR-Cas9 systems for genomic editing, regulation and targeting. Nat. Biotechnol. 32, 347-55 (2014).
 
■ 产品详情请点击:Human iPS Cell Editing Systems
 
 
 
Human iPS Cell Editing Systems
Human induced pluripotent stem cells (hiPSCs) growing in DEF-CS medium were edited with Cas9 and sgRNA specific to CD81. Pluripotency is maintained in edited human induced pluripotent stem cells (hiPSCs) that have been seeded as single cells. 12 clonal lines created from single-seeded hiPSCs exhibit high levels of Oct-4 (97-99%), TRA-1-60 (98-99%), and SSEA-4 (98-99%).
  (图片来源于Takara Bio USA, Inc.)
 
 
 

13201 | PKH26 Cell Membrane Labeling Kit, 100 uL dye, – Lumiprobe荧光染料

上海金畔生物科技有限公司代理lumiprobe荧光染料全线产品,欢迎访问官网了解更多信息

13201 | PKH26 Cell Membrane Labeling Kit, 100 uL dye,

¥2,470.00

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SKU: Lumiprobe.13201 分类: 默认分类

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Human iPS Cell Generation™ All-in-One Vector酶试剂盒Takara Clontech

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Human iPS Cell Generation All-in-One Vector
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Takara 3671 Human iPS Cell Generation All-in-One Vector 10 μg ¥10,014 Human iPS Cell Generation™ All-in-One Vector Human iPS Cell Generation™ All-in-One Vector Human iPS Cell Generation™ All-in-One Vector
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□ 浓度 0.5 μg/μl
     
□ 贮存溶液  
     Tris-HCl (pH 8.0) 10 mM
     EDTA 1 mM
Limited Use Label License: [L44]
 
■ 制品说明
本制品是制备Human iPS细胞诱导用逆转录病毒所使用的载体。在高效、高表达逆转录病毒载体pDON-5 DNA(Code No.: 3658)上,插入了iPS细胞诱导用基因OCT3/4SOX2KLF4LIN28NANOG,各基因以Thosea asigna virus 的2A*1序列连接在一个载体上。本载体与Retrovirus Packaging Kit Ampho(Code No.: 6161)中的pGP Vector以及pE-ampho Vector共转染至逆转录病毒制备用细胞G3T-hi(Code No.: 6163)后,可以制备iPS诱导用逆转录病毒。如此制备的重组逆转录病毒由于和RetroNectin®(Recombinant Human Fibronectin Fragment)(Code No.: T100A/B)有高亲和性,所以在向目的细胞中导入基因时,使用经RetroNectin®处理后的培养皿或平板,可以得到较高的基因导入率,有效提高iPS细胞的诱导效率。
*1 5种基因转录在同一个mRNA上,翻译时在2A序列的特定位点被分割,使5种蛋白质可以等量表达。
 
■ 保存
-20
 
■ 各载体的遗传信息
  基因名   GenBank Accession No.
  OCT4   NM_002701.4
  SOX2   NM_003106.2
  KLF4   NM_004235.3*2
  LIN28   NM_024674.4
  NANOG   NM_024865.2
*2 更新的NM_004235.4中,CDS的5’端追加了对应9个氨基酸的碱基。本制品的KLF4基因中不含N端的这9个氨基酸的序列,已经确认可以有效诱导iPS细胞。
 
■ 载体图谱
Human iPS Cell Generation™ All-in-One Vector
 
 
 
 

 

23201 | PKH26 Cell Membrane Labeling Kit, 100 uL dye, – Lumiprobe荧光染料

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23201 | PKH26 Cell Membrane Labeling Kit, 100 uL dye,

¥1,820.00

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SKU: Lumiprobe.23201 分类: 默认分类

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Cell Lysis Buffers酶试剂盒Takara Clontech

上海金畔生物科技有限公司代理Takara Clontech酶试剂盒全线产品,欢迎访问官网了解更多产品信息和订购。

Cell Lysis Buffers
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Clontech 635625 TALON xTractor Buffer 500 ml ¥4,422 Cell Lysis Buffers Cell Lysis Buffers Cell Lysis Buffers
Clontech 635656 xTractor Buffer 100 ml ¥1,616 Cell Lysis Buffers Cell Lysis Buffers Cell Lysis Buffers
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Code No. 产品名称
635625   xTractor Buffer
635656   xTractor Buffer
635671   xTractor Buffer
635623   xTractor Buffer Kit
 
√    通用 可从细菌、酵母、哺乳动物细胞和杆状病毒感染的细胞中提取蛋白质
√    高效率 效果优于sonic或其他细胞裂解液
√    快速 流程简便,只需10min孵育
√    温和 非变性提取条件,更有利于保持生物活性
√    兼容 适用于所有IMAC树脂,快速纯化标签蛋白质
 
Cell Lysis Buffers
快速纯化His标签蛋白操作流程图
 
产品详情请点击:Cell Lysis Buffers
 
 

Chaperone Competent Cell BL21 Series酶试剂盒Takara Clontech

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Chaperone Competent Cell BL21 Series
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Takara 9120 Chaperone Competent Cells BL21 Set 1 Set ¥2,613 Chaperone Competent Cell BL21 Series Chaperone Competent Cell BL21 Series Chaperone Competent Cell BL21 Series
Takara 9121 Chaperone Competent Cell pG-KJE8/BL21 100 μl × 10 ¥764 Chaperone Competent Cell BL21 Series Chaperone Competent Cell BL21 Series Chaperone Competent Cell BL21 Series
Takara 9122 Chaperone Competent Cell pGro7/BL21 100 μl × 10 ¥535 Chaperone Competent Cell BL21 Series Chaperone Competent Cell BL21 Series Chaperone Competent Cell BL21 Series
Takara 9124 Chaperone Competent Cell pG-Tf2/BL21 100 μl × 10 ¥1,106 Chaperone Competent Cell BL21 Series Chaperone Competent Cell BL21 Series Chaperone Competent Cell BL21 Series
Takara 9125 Chaperone Competent Cell pTf16/BL21 100 μl × 10 ¥1,232 Chaperone Competent Cell BL21 Series Chaperone Competent Cell BL21 Series Chaperone Competent Cell BL21 Series
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■ 制品内容Chaperone Competent Cell BL21 Series(Code No.: 9120:100 μl × 3 × 6 种)
● Chaperone Competent Cells BL21 Set (Code No.: 9120)
1. Chaperone Competent Cells pG-KJE8/BL21 3 × 100 μl
2. Chaperone Competent Cells pGro7/BL21 3 × 100 μl
3. Chaperone Competent Cells pKJE7/BL21 3 × 100 μl
4. Chaperone Competent Cells pG-Tf2/BL21 3 × 100 μl
5. Chaperone Competent Cells pTf16/BL21 3 × 100 μl
6. TaKaRa Competent Cell BL21 3 × 100 μl
7. pUC19 DNA (0.1 ng/μl) 1× 10 μl
8. SOC medium* 18 × 1 ml
   
● Chaperone Competent Cells pG-KJE8/BL21 (Code No.: 9121)
● Chaperone Competent Cells pGro7/BL21 (Code No.: 9122)
● Chaperone Competent Cells pG-Tf2/BL21 (Code No.: 9124)
● Chaperone Competent Cells pTf16/BL21 (Code No.: 9125)
1. Competent Cell 10 × 100 μl
2. pUC19 DNA (0.1 ng/μl) 1× 10 μl
3. SOC medium* 10 × 1 ml
*SOC medium:       2%   Tryptone
 0.5%   Yeast extract
 10 mM   NaCl
 2.5 mM   KCl
 10 mM   MgSO4
 10 mM   MgCl2
 20 mM   Glucose
     
 
■ 制品说明
Chaperone Competent Cell BL21 Series是由五种分别转化一种伴侣蛋白质粒 (Chaperone Plasmid Set (Code No.: 3340)) 的Escerichia coli BL21制成的感受态细胞。
众所周知,分子伴侣参与蛋白质折叠过程,且人们已经通过大量的实验研究来表明体内蛋白折叠机理。 本制品中含有五种类型的质粒(pG-KJE8、pGro7、pKJE7、pG-Tf2、pTf16),每一种质粒都是能有效表达一组协同作用、共同参与蛋白折叠的被称作“Chaperone team”的分子伴侣。靶蛋白与其中一种“Chaperone team”共同表达,便可增加可溶性蛋白质的回收比率,而用通常方法,由于包涵体的形成,这些表达蛋白质常常是不可回收的。
E.coli BL21来源于B株,为Lon蛋白水解酶、ompT外膜蛋白水解酶缺陷型。因此,表达的外源蛋白质在该体系中有更高的稳定性。
当进行靶蛋白与“Chaperone team”共表达实验时,通常需要三步:
1. 用伴侣蛋白质粒转化宿主E.coli; 2. 用转化体制备感受态细胞;3. 用质粒转化感受态细胞并表达靶蛋白。如果使用本产品,只需一步转化就可构建靶蛋白与“Chaperone team”的共表达体系,因为使用的E.coli BL21感受态细胞已转化有一种伴侣蛋白质粒。另外,此感受态细胞有五种质粒,不含有伴侣蛋白质粒的TaKaRa Competent BL21也可作为对照。
每种E.coli BL21感受态细胞包括一种伴侣蛋白质粒,非常适合pCold DNA系列的表达。但该产品不适合于启动子为T7的蛋白质表达体系,如pET系列,因为,宿主E.coli BL21菌株不表达T7 RNA聚合酶。
注意:本产品不可用于电击法转化。
 
■ DNA序列
  pG-KJE8 DNA (Fasta形式的txt文件)
  pGRO7 DNA (Fasta形式的txt文件)
  pG-TF2 DNA (Fasta形式的txt文件)
  pKJE7 DNA (Fasta形式的txt文件)
  pTf16 DNA (Fasta形式的txt文件)
 
■ 保存
-80℃。
注意:如果不是-80℃保存,转化效率将会降低。此时,可在使用前用附带的pUC19做转化实验,确认转化效率。不能保存在液氮中。
 
■ 转化效率
Chaperone competent cells:转化1 ng pUC19,涂于Cm+和Amp+平板上进行筛选。
TaKaRa competent cell BL21:转化1 ng pUC19,涂于Amp+平板上进行筛选。
转化效率≥1×106 cfu/μg pUC19
 
■ Genotype
E.coli BL21 : F, ompT, hsdSB (rBmB), gal, dcm
 
■ 注意事项
1. 从-80℃中只取出实验所用的感受态细胞,然后立即放入干冰/EtOH bath。开始实验前不要取出。
2. 可使用1.5 ml microtube替代FALCON tube,但可能会降低效率。
3. 使用100 μl感受态细胞转化时,高纯度DNA使用量不要超过10 ng,否则转化效率会降低。
4. 转化体系改变 (如感受态细胞使用量发生改变) ,或换用了其它tube等,需研讨转化条件 (如使用一般的1.5 ml tube时,请于42℃保温60秒)。
5. 除SOC培养基外,L-broth及φb-broth也可使用,但转化效率会有所下降。
6. 稀释时,使用SOC培养基。
7. L-broth plate : 成份 per liter  
  Tryptone 10 g  
  Yeast extract 5 g  
  NaCl 5 g  
    用1N NaOH调整到pH7.5,添加agar到1.5%,高压灭菌。
8. 不建议将解冻的感受态细胞再次冷冻保存。如果再次冻结不可避免,将细胞置于干冰/乙醇中迅速冷冻,置于-80℃保存。但是,转化效率可能会降低至少一个数量级。
9. 本制品中含有的伴侣蛋白质粒带有来源于pACYC的复制起点和氯霉素抗性基因 (Cmr),E.coli表达系统中可以使用常用的ColE1型、具氨苄抗性 (Ampr)筛选标记的表达载体,但不可与含有氯霉素抗性基因的表达质粒共同使用。