牛胶原蛋白溶液Type I, 3 mg/ml
英文名：PureCol®, Bovine Collagen, 3 mg/mL, 100 mL
货号：BM-5005-100ML
规格：100 ml
品牌： Advanced Biomatrix
报价：￥5800.00
商品描述
PureCol® collagen is known as the standard of all collagens for purity (>99.9% collagen content), functionality, and the most native-like collagen available. PureCol® is isolated from bovine hides sourced from the only controlled, closed herd in the United States. Advanced BioMatrix’s manufacturing processes comply with stringent quality standards that have proven to yield unsurpassed lot-to-lot consistency.
PureCol® collagen is approximately 97% Type I collagen with the remainder being comprised of Type III collagen. It contains a high monomer content as measured by gel permeation chromatography. PureCol® collagen is supplied at approximately a 3 mg/ml concentration. The concentration for each specific lot is provided on a Certificate of Analysis that is available with the purchase of each product. PureCol® is soluble atelo-collagen in 0.01 N HCI, therefore, the pH is approximately 2.0. PureCol® collagen is ideal for coating of surfaces, providing preparation of thin layers for culturing cells, or use as a solid gel.
PureCol® collagen is provided in a user-friendly packaging for use and storage. PureCol® is sterile filtered and is supplied as a ready to use solution.
Purecol胶原蛋白被称为标准纯度的胶原蛋白（>99.9%胶原蛋白含量），其功能可媲美大多数的原生胶原蛋白。Purecol提取于来自美国封闭式牛群的牛皮，先进的生物生产过程符合严格的质量标准，已被证明是质量超凡的。Purecol胶原蛋白是由约97%的I型胶原蛋白，其余的为III型胶原蛋白组成的，它具有很高的单体含量可以用于测试凝胶渗透色谱。本产品的浓度约为3mg/ml，每个批次的具体浓度会写于产品的COA上随货发出。PureCol是溶于0.01N盐酸的端胶原蛋白，因此它的PH值约为2。PureCol胶原蛋白是理想的蛋白质涂层，可用于准备培养的细胞或者固体凝胶。PureCol胶原蛋白的包装便于储存和使用，经过灭菌的溶液随时可以使用。
如何制作 3D 胶原蛋白水凝胶
如何使用 I 型胶原蛋白有效地形成 3D 胶原蛋白凝胶。 成功的胶原蛋白水凝胶的 4 个主要要求是：
1. 中性 pH (7.0-7.4)
2. 等渗盐 (~300 mOsmo)
3. 温度（37°C）
4. 浓度 (>1.5 mg/ml)
Collagen Gelation Diagnosis
Reasons Your Collagen Might Not be Gelling
The pH is not neutral
Double check to make sure the pH is 7.0-7.4.
Even if using a neutralization solution, the pH may be off due to mixing issues or collagen "hold-up" in pipette tips, slightly altering the collagen:neutralization solution ratio.
Incorrect levels of salts
Ensure the salts of the neutralization collagen solution are isotonic (~300 mOsmo). We accomplish this by adding 1 part of 10X PBS or Media in most collagen hydrogel preparations.
Temperature
Collagen gels best at 37C. Ensure the temperature does not exceed 38.5C, as the collagen will begin to denature.
Collagen Concentration
Low collagen concentration solutions will have a hard time forming a gel. We recommend keeping the collagen concentration >1.5 mg/ml for best results.
Mechanical agitation
Vortexing the collagen solution can have negative impacts on the collagen and will negatively affect the ability of the solution to form a hydrogel. Do not vortex.
Do not agitate the collagen solution during incubation and gelation. This will interfere with the fibrillogenesis process.
Material was frozen or left at room temperature
If the collagen solution was frozen or left at room temperature for a long period of time, it may have begun to denature, which would affect its ability to gel.
Very small volumes of collagen were used in a non-confined space
We have found that using microliters of collagen on top of glass coverslips tend to not gel very well.
We recommend ensuring the collagen is within a confined space (ie. wells or walls)
Neutralization Solution came out of solution
If using a collagen product that comes with a neutralization solution, ensure that the neutralization solution is still fully in solution. Getting it too cold can cause it to come out of solution. If this happens, simply warm it back up.
Weights vs Volume
Due to pipette hold-up volume, dispensing 1 mL of collagen (3 mg/ml) using a pipettor actually only dispenses about .89 grams. For the most accurate results, we recommend usings weights for viscous materials, such as collagen. The neutralization solutions, buffers and medias can use volume, as their viscosity is close to water.
Expired material
Collagen tends to be stable when stored properly, but after a few years, the "gel strength" slowly declines. Expired collagen may still form hydrogels, but it will be significantly softer than a fresh batch of collagen.
Heat transfer
Warming collagen to 37C for polymerization in a test tube, in a water batch will be much more efficient than gelling collagen in an enclosed 6-well plate, with a lid, in a dry incubator. Look for ways to improve heat transfer if quicker polymerization is desired.
Mixing
Ensure proper mixing to create a homogeneous solution (without vortexing) for best results.
Lyophilized Collagen
Atelocollagen that has been lyophilized (such as lyophilized PureCol, catalog #5006), tends to form weaker hydrogels, or not polymerize at all.
Insoluble Collagen
Advanced BioMatrix offers two insoluble collagen products, #5162 and #5164. These collagen powders are extremely robust and will not fully go into solution.
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产品信息
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PureCol® 胶原蛋白溶液
货号 # BM-5005-100ML |
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灭菌方法
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过滤
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形态
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溶液
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包装尺寸
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100 ml
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储存温度
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2-10 °C
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有效期
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详见产品包装与随货发出的COA
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浓度（缩二脲蛋白测定）
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2.9-3.2 mg/ml
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纯度（胶原蛋白浓度）
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> 99.9%
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pH
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pH约为2, 0.01 N HCl
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凝胶时间（凝胶时间测定法）
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< 40 分钟
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原纤维形成（纤维形成）
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>0.5 吸光度单位
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聚丙烯酰胺凝胶电泳（SDS–PAGE）
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> 85%含胶原蛋白在α，β，和γ带，< 15%含比α带快的胶原蛋白
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无菌的
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没有细菌生长
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内毒素 (LAL)
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< 1.0 EU/ml
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渗透压
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~ 20 mOsm
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电导率
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~ 2300 µs/cm
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细胞附着实验
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通过
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来源
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牛皮–胃蛋白酶提取
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使用方法：
Coating Procedure
Note:Use these recommendations as guidelines to determine the optimal coating conditions for your culture system.
- Remove required quantity of collagen from the bottle and dispense into a dilution vessel.
- Dilute PureCol® in water to ~50 to 100 µg/ml (~1:30). A 0.01 M HCl solution may also be used.
- Swirl contents gently until material is completely mixed.
- Add appropriate amount of diluted PureCol® material to the culture surface ensuring that the entire surface is coated.
- Incubate at room temperature or 37°C, covered, for 1-2 hours.
- After incubation, aspirate any remaining material.
- Rinse coated surfaces carefully with sterile medium or PBS, avoid scratching surfaces.
- Coated surfaces are ready for use. They may also be stored at 2-8°C damp or air dried if sterility is maintained.
3-D Gel Preparation Procedure
- Slowly add 1 part of chilled 10X PBS or 10X culture media to 8 parts of chilled collagen solution with gentle swirling.
- Adjust pH of mixture to 7.2–7.6 using sterile 0.1 M NaOH. Monitor pH adjustment carefully (pH meter, phenol red, or pH paper).
- Adjust final volume to a total of 10 parts with sterile water.
- To prevent gelation, maintain temperature of mixture at 2–10°C.
- To form gel, warm to 37°C. Allow approximately 90 to 120 minutes for gel formation.
Cell Attachment Bioassay
To demonstrate cell attachment, cells were seeded onto surfaces coated with the matrix product and onto negative control surfaces without the matrix product. Cells were seeded onto surfaces using serum-free DMEM media. All surfaces were blocked with a solution containing 1% BSA. Cells were then allowed to attach for one (1) hour at 37°C. The culture surfaces were washed with DPBS. The results indicate significant cell attachment with surfaces coated using the matrix product.
| Without UltraPure® | With UltraPure® |
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| Human Umbilical Vein Endothelial Cells (HUVEC) 10X | |
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| VERO Epithelial Cells 10X | |
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| Human Dermal Fibroblast Cells 10X | |
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| MDCK Epithelial Cells 10X | |
注意事项：
This product is for R&D use only and is not intended for human or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.
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