重组人MEGACD40L® 蛋白(可溶)


产品编号 产品名称 产品规格 产品等级 产品价格
ALX-522-110-C010  MEGACD40L (soluble) (human), (recombinant)
重组人Mega CD40L(可溶) 
10 μg
  • 产品特性
  • 相关资料
  • Q&A
  • 参考文献

重组人MEGACD40L® 蛋白(可溶)重组人MEGACD40L® 蛋白(可溶)

用于免疫反应中协同刺激激活的高活性、高纯度CD40L蛋白质

◆特点

● 高活性MEGACD40L® 低聚物模拟体内膜辅助CD40L聚合和激活而无需增强剂

● 高纯度,低内毒素水平

● 丰富的生产经验保证产品批次间一致性和可靠性

● 优化的配方以增强稳定性并提高活性

MEGACD40L是一种高活性蛋白,是两个三聚体CD40配体分子通过Adiponectin/ACRP30/AdipoQ的胶原域人工连接而成。在体内,这种蛋白质能有效的模拟CD40L的天然膜辅助聚合。可简单有效的替代 [CD40L+增强剂]组合(产品编号:ALX-850-064)。

◆产品规格

别名

  CD40L:ACRP30headless, CD154:ACRP30headless,

  TNFSF 5:ACRP30headless,  gp39:ACRP30headless

分子量

  ~38 kDa

来源

  CHO细胞表达。人CD40L(CD154)(aa 116-261)与小鼠ACRP30headless

  (aa 18-111)N端以及一个FLAG® -tag融合。

UniProt ID

  P29965 (human CD40L), Q60994 (mouse Adiponectin)

浓度

  复溶后0.1 mg/mL

产品类型

  冻干,含PBS

纯度

  ≥90%(SDS-PAGE)

纯化细节

  多步层析纯化

内毒素含量

  ≤0.01EU/μg 纯化蛋白(LAL 测试;Cape Cod Associates)

生物活性

  与人CD40结合。B细胞激活(已证明存在CD86剂量依赖性上调)

应用

  可用于调节免疫和炎症反应,作为免疫治疗的临床前研究的一种蛋白质工具,

  用来监测病人接受化疗的过程以及模拟自体免疫反应表型。

配制

  用100 μL的无菌水溶解。以含5%胎牛血清的培养基进行进一步稀释。

运输

  冰袋运输

长期储存

  -20℃

使用/稳定性

  收货后可稳定储存在-20℃至少6个月

处理

  避免反复冻融。复溶后,分装成小份储存于-20℃。

技术信息/产品说明

  FLAG是Sigma-Aldrich Co.的注册商标。

◆应用实例

重组人MEGACD40L® 蛋白(可溶)

MEGACD40L® (产品编号:ALX-522-110)激活B淋巴细胞。方法:外周血单核细胞(PBMCs)37℃、5%CO2 在48孔板(1×106 细胞/孔)中培养48小时。每孔包含200 μL已梯度稀释的MEGACD40L® 无血清测试培养基。处理后,清洗细胞,用小鼠anti-human CD19-PE和小鼠anti-humanCD86-APC进行双重染色,然后用流式细胞术进行分析。所示数据为在不同的MEGACD40L® 浓度下,共染色为CD19+ 和CD86+ B细胞的百分数。

重组人MEGACD40L® 蛋白(可溶)

重组人MEGACD40L®(可溶)(产品编号:ALX-522-110)结构示意图


重组人MEGACD40L® 蛋白(可溶)

PBMCs(外周血单核细胞)37℃、5%CO2 在48孔板(1×106细胞/孔)中培养48小时,每孔包含200 μL已梯度稀释的MEGACD40L®(产品编号:ALX-522-110)无血清测试培养基、CD40L(产品编号:ALX-522-015)和CD40L+2 μg/mL 增强剂(产品编号:ALX-804-034)。处理后,用PBS+1%BSA清洗细胞3次。用小鼠anti-human CD19(PE偶联)和小鼠anti-humanCD86(APC偶联)进行双重染色,4℃,2小时。用PBS+1%BSA清洗细胞3次,用PBS重悬。样品用BD Facs Calibur 流式细胞分析仪进行分析。所示数据为在不同的浓度下CD86+B细胞百分数/CD19+B细胞百分数。

重组人MEGACD40L® 蛋白(可溶)

1000 ng/mL、7个连续批次的MEGACD40L® 用于B细胞激活实验数据


重组人MEGACD40L® 蛋白(可溶)

通过在指定的温度下储存蛋白2个月来进行MEGACD40L® 稳定性测试。用B细胞激活实验来进行蛋白活性测试。结果显示在严苛的储存条件下MEGACD40L® 仍能保持较好的活性。

◆相关产品


产品编号

产品名称

包装

产品描述

ALX-522-120-C010

MEGACD40L (soluble) (mouse), (recombinant)
MegaCD40L (可溶性) (小鼠), (重组)

10 μg

用于免疫反应中协同激活活性的高活性、高纯度CD40L蛋白。

ALX-850-064-KI01

CD40L, soluble (human) (recombinant) set
CD40L, 可溶性 (人) (重组) 套装

1 set

配体+增强剂用于提高稳定性和增强免疫活性。

ADI-CSA-187-E

CD 40L monoclonal antibody (5F3)
CD40L, 单抗 (5F3)

100 μg

流式细胞术

ALX-805-037-C100

CD40L (human) monoclonal antibody (blocking) (MK13A4)
CD40L (人), 单抗 (封闭作用) (MK13A4)

100 μg

流式细胞术,FUNC

ALX-522-015-6010

CD40L (soluble) (human), (recombinant)
  CD40L (可溶性) (人), (重组)

6×10 μg

增加对B细胞的刺激。
E.coli表达。人CD40L(CD154)(aa 116-261)胞外区域与一个短肽(6   aa)的N端以及一个FLAG®-tag融合,≥95%(SDS-PAGE),ELISA

ALX-522-015-2010

2×10 μg

ALX-522-015-C010

10 μg

参考文献

重组人MEGACD40L® 蛋白(可溶)


MEGACD40L-cited-examples

 1.

Artesunate shows potent anti-tumor activity in B-cell lymphoma: T. K. Vatsveen, et al.; J. Hematol. Oncol. 11, 23 (2018)


 2.

CD40L and TNF both activate the classical NF-κB pathway, which is not required for the CD40L induced alternative pathway in endothelial cells: J. Seigner, et al.; Biochem. Biophys. Res. Commun. 495, 1389 (2018)


 3.

CD40L mediated alternative NFκB-signaling induces resistance to BCR-inhibitors in patients with mantle cell lymphoma: H. Rauert-Wunderlich, et al.; Cell Death Dis. 9, 86 (2018)


 4.

Characterisation of anifrolumab, a fully human anti-interferon receptor antagonist antibody for the treatment of systemic lupus erythematosus: J.M. Riggs, et al.; Lupus Sci. Med. 5, (2018)


 5.

Engineering protein-secreting plasma cells by homology-directed repair in primary human B cells: K.L. Hung, et al.; Mol. Ther. 26, 456 (2018)


 6.

High PD-L1/CD86 MFI ratio and IL-10 secretion characterize human regulatory dendritic cells generated for clinical testing in organ transplantation: A.F. Zahorchak, et al.; Cell. Immunol. 323, 9 (2018)


 7.

In vitro antineoplastic effects of auranofin in canine lymphoma cells: H. Zhang, et al.; BMC Cancer 18, 522 (2018)


 8.

CD40 signaling in Graves disease is mediated through canonical and noncanonical thyroidal nuclear factor κB activation: H.J. Lee, et al.; Endocrinology 158, 410 (2017)


 9.

Cutting Edge: Active TGF-β1 Released from GARP/TGF-β1 Complexes on the Surface of Stimulated Human B Lymphocytes Increases Class-Switch Recombination and Production of IgA: O. Dedobbeleer, et al.; J. Immunol. 199, 391 (2017)


10.

Down‐regulation of CD73 on B cells of patients with viremic HIV correlates with B cell activation and disease progression: E.S. Kim, et al.; J. Leukoc. Biol. 101, 1263 (2017)


11.

Human secretory IgM emerges from plasma cells clonally related to gut memory B cells and targets highly diverse commensals: G. Magri, et al.; Immunity 47, 118 (2017)


12.

Regulation of cell surface CB2 receptor during Human B cell activation and differentiation: J.T. Castaneda, et al.; J. Neuroimmune Pharmacol. 12, 544 (2017)


13.

CD40 expression in fibrocytes Is induced by TSH: potential synergistic immune activation: T. Mester, et al.; PLoS One 11, e0162994 (2016)


14.

Distinct TLR-mediated cytokine production and immunoglobulin secretion in human newborn naïve B cells: M.A. Pettengill, et al.; Innate Immun. 22, 433 (2016)


15.

Expression of HIV-1 matrix protein p17 and association with B-cell lymphoma in HIV-1 transgenic mice: V.A. Carroll, et al.; PNAS 113, 13168 (2016), Application(s): Human B cells culture


16.

T-helper signals restore B-cell receptor signaling in autoreactive anergic B cells by upregulating CD45 phosphatase activity: P. Szodoray, et al.; J. Allergy Clin. Immunol. 138, 839 (2016), Application(s): Cell culture


17.

A human monoclonal antibody targeting the conserved staphylococcal antigen IsaA protects mice against Staphylococcus aureus bacteremia: S. van den Berg, et al.; Int. J. Med. Microbiol. 305, 55 (2015), Application(s): ELISA, PCR of human blood samples


18.

CD40L Induces Functional Tunneling Nanotube Networks Exclusively in Dendritic Cells Programmed by Mediators of Type 1 Immunity: C. Zaccard, et al.; J. Immunol. 194, 1047 (2015), Application(s): Activation of mature human dendritic cells


19.

Detection of IκB degradation dynamics and IκB-α ubiquitination: P. Starokadomskyy, et al.; Methods Mol. Biol. 1280, 15 (2015)


20.

Establishing the pig as a large animal model for vaccine development against human cancer: N.H. Overgaard, et al.; Front. Genet. 6, 286 (2015), Abstract; Full Text


21.

Haploinsufficiency of the NF-κB1 subunit p50 in common variable immunodeficiency: M. Fliegauf, et al.; Am. J. Hum. Genet. 97, 389 (2015)


22.

Mechanism and functional impact of CD40 ligand-induced von Willebrand factor release from endothelial cells: K. Möller, et al.; Thromb. Haemost. 113, 1095 (2015)


23.

NF-κB, p38 MAPK, ERK1/2, mTOR, STAT3 and increased glycolysis regulate stability of paricalcitol/dexamethasone-generated tolerogenic dendritic cells in the inflammatory environment: K. Danova, et al.; Oncotarget 6, 14123 (2015)


24.

Phenotypic Approaches to Identify Inhibitors of B Cell Activation: E. B. Rex, et al.; J. Biomol. Screen. 20, 876 (2015), Application(s): Assay


25.

Systemic agonistic anti-CD40 treatment of tumor-bearing mice modulates hepatic myeloid-suppressive cells and causes immune-mediated liver damage: J. Medina-Echeverz, et al.; Cancer Immunol. Res. 3, 557 (2015)


26.

A GPBAR1 (TGR5) small molecule agonist shows specific inhibitory effects on myeloid cell activation in vitro and reduces experimental autoimmune encephalitis (EAE) in vivo: N. Lewis, et al.; PLoS One 9, e100883 (2014), Application(s): Assay of human PBMC


27.

Development of a novel anti-canine CD20 monoclonal antibody with diagnostic and therapeutic potential: D. Ito, et al.; Leuk. Lymphoma 56, 219 (2014), Application(s): Analysis of canine B-cell lymphoma cells


28.

Exosomes Derived from Burkitt’s Lymphoma Cell Lines Induce Proliferation, Differentiation, and Class-Switch Recombination in B Cells: C. Gutzeit, et al.; J. Immunol. 192, 5852 (2014), Application(s): PCR, Assay of human B cells


29.

Preclinical evaluation of the novel, orally bioavailable Selective Inhibitor of Nuclear Export (SINE) KPT-335 in spontaneous canine cancer: results of a phase I study: C. London, et al.; PLoS One 9, e87585 (2014), Application(s): Analysis of canine tumor cells by WB, FLOW


30.

Thyrotropin receptor and CD40 mediate interleukin-8 expression in fibrocytes: implications for thyroid-associated ophthalmopathy (an American Ophthalmological Society thesis).: R. Douglas, et al.; Trans. Am. Ophthalmol. Soc. 12, 26 (2014), Application(s): In vitro analysis of human fibrocytes


31.

AMPKα1 deficiency amplifies proinflammatory myeloid APC activity and CD40 signaling: K.C. Carroll, et al.; J. Leukoc. Biol. 94, 1113 (2013)


32.

Blood CD4+CD45RO+CXCR5+ T cells are decreased but partially functional in signal transducer and activator of transcription 3 deficiency: F. Mazerolles, et al.; J. Allergy Clin. Immunol. 131, 1146 (2013), Application(s): Assay of blood cells


33.

CD40 ligand is necessary and suffi cient to support primary diff use large B-cell lymphoma cells in culture: a tool for in vitro preclinical studies with primary B-cell malignancies: D. Ito, et al.; Leuk. Lymphoma 53, 1390 (2012), Application(s): Flow cytometry of human B cells


34.

HIV-1 gp120 impairs the induction of B cell responses by TLR9-activated plasmacytoid dendritic cells: N.P. Chung, et al.; J. Immunol. 189, 5257 (2012), Application(s): Activation of dendritic cells


35.

Immunodeficiency, autoinflammation and amylopectinosis in humans with inherited HOIL-1 and LUBAC deficiency: B. Boisson, et al.; Nat. Immunol. 13, 1178 (2012), Application(s): ELISA of human blood cells


36.

Interleukin-6 Production in CD40-Engaged Fibrocytes in Thyroid-Associated Ophthalmopathy: Involvement of Akt and NF-κB: E.F. Gillespie, et al.; Invest. Ophthalmol. Vis. Sci. 53, 7746 (2012), Application(s): CD40 ligation, induction and signaling in Graves' orbital fibroblasts (GOFB) and fibrocytes


37.

Siglec-7 tetramers characterize B-cell subpopulations and leukemic blasts: F. Gieseke, et al.; Eur. J. Immunol. 42, 2176 (2012), Application(s): ELISA of human B cells


38.

New mechanism of X-linked anhidrotic ectodermal dysplasia with immunodeficiency: impairment of ubiquitin binding despite normal folding of NEMO protein: M. Hubeau, et al.; Blood 118, 926 (2011)


39.

Reconstitution of huPBL-NSG mice with donor-matched dendritic cells enables antigen-specific T-cell activation: A. Harui, et al.; J. Neuroimmune Pharmacol. 6, 148 (2011), Application(s): Activation of dendritic cells


40.

Study of patients with Hyper-IgM type IV phenotype who recovered spontaneously during late childhood and review of the literature: N. Karaca, et al.; Eur. J. Pediatr. 170, 1039 (2011), Application(s): Determining human B-cell function in Hyper-IgM blood samples


41.

Human TRAF3 adaptor molecule deficiency leads to impaired Toll-like Receptor 3 response and susceptibility to Herpes Simplex Encephalitis: R. Perez de Diego, et al.; Immunity 33, 400 (2010)


42.

Multimeric soluble CD40 ligand (sCD40L) efficiently enhances HIV specific cellular immune responses during DNA prime and boost with attenuated poxvirus ventors MVA and NYVAC expressing HIV antigens : C.E. Gomez, et al.; Vaccine 27, 3165 (2009)


43.

Impaired CD40L signaling is a cause of defective IL-12 and TNF-alpha production in Sezary syndrome: circumvention by hexameric soluble CD40L: L.E. French, et al.; Blood 105, 219 (2005)


44.

Two adjacent trimeric Fas ligands are required for Fas signaling and formation of a death-inducing signaling complex: N. Holler, et al.; Mol. Cell. Biol. 23, 1428 (2003)

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