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查看更多产品信息 Collagen I, rat tail - FAQs (A1048301)
19 个常见问题解答
分子量为300 kDa。
I型胶原是以20mM乙酸溶液的液体形式来提供的。如果在准备过程中遇到困难，可先按1:2的比例将胶原稀释于20mM，pH 3.5的乙酸中，之后再按所用的实验方案中的说明进一步稀释。
可用的酶包括：Gibco TrypLE Express，Gibco TrypLE Select或Gibco StemPro Accutase 细胞解离试剂（Cell Dissociation Reagent）。
以下细胞类型通常培养于I型胶原中：
- 内皮细胞（包括HUVEC）
- 成纤维细胞
- 肝细胞
- 软骨细胞
- 破骨细胞
- 成骨细胞
- 角质形成细胞
- 角膜上皮
其关键应用列举如下：
- I型胶原能够在体外诱导微血管内皮细胞形成纺锤状形态，并排列为类似实线的聚集体。在I型胶原上进行培养时，血管内皮细胞也能够形成类似血管的管状结构。因此，I型胶原也可用于体外的血管生成实验。
- 乳腺癌干细胞在I型胶原上培养时可发生分化。
- I型胶原同样已被用于原代结肠癌细胞系的培养；小鼠肝脏前体细胞和大鼠的胰岛细胞也可培养于3D的I型胶原中。
- I型胶原可在细胞侵润实验分析中作为屏障，也已被用于细胞粘附的相关研究。
Gibco Geltrex 基质与胶原（大鼠尾与牛）可作为包被溶液或3D凝胶基质来使用。CELLstart 底物作为一种不含异外源性物质的包被基质，专为ESC的相关应用而研发（作为动物源性的Gibco Geltrex 基质或动物源性的Matrigel 基质的一种替代品底物）。AlgiMatrix 基质是为一种3D支架类的基质，它不支持并不为细胞提供粘附的基础，但而是为细胞球生长提供一个良好的生长环境，后续应用中也可方便易于细胞球的收取操作用于后续应用。
The polystyrene that culture flasks are made of tends to have a negative charge. Many cell types will still attach because the plasticware is treated in a way to make them hydrophilic. However cell types that do not attach well to the plastic, or are grown in medium with little or no serum protein, will attach better if the plastic is coated with a protein/polymer which has an overall positive charge.
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No, the Collagen I, rat tail is not pepsin-digested.
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Unfortunately, we do not have an animal-origin-free alternative for the Coating Matrix Kit Protein (Cat. No. R011K).However, we offer Rat or Bovine collagen that can be used as coating matrices. Please see the product pages linked below:
Collagen I, bovine (Cat. No. A1064401)
Collagen I, rat tail (Cat. No. A1048301)
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An alternative for Collagen I, rat tail (Cat. No A1048301) is Collagen I, bovine (Cat. No A1064401). While the source of Collagen I is different, they can both be prepared as a clear gel providing a 3D matrix or surface coating on tissue culture plates as a substrate for culturing primary cells such as keratinocytes and hepatocytes.
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You can just use water to dilute the acetic acid stock solution to 20 mM.
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For your application, the thin coating procedure should be fine at a starting concentration of 5 µg/cm2. You can dilute the collagen in 20 mM acetic acid to the volume needed. Coat the surface, incubate 1 hour at room temp, aspirate the solution, rinse 3 times with PBS, and use or store plates.
See the manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/Collagen_I_rat_tail_PI.pdf) for details.
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The pH of the diluted rat tail collagen will determine whether the rat tail collagen is a liquid or a gel. A pH of 5 or below will not form any gel. If the pH is above 8, the collagen will not form a gel but will precipitate in small particles. A good gel can be formed at pH between 6.5 and 7.5.
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The molecular weight is 300 kDa.
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Collagen I is provided as a liquid, suspended in 20 mM acetic acid solution. If difficulties are encountered with the preparation, the collagen can be diluted in 1:2, first in 20 mM acetic acid, pH 3.5 before further diluting according to the protocol being used.
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Enzymes that can be used include: Gibco TrypLE Express Enzyme, Gibco TrypLE Select Enzyme, or Gibco StemPro Accutase Cell Dissociation Reagent.
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The following cell types are commonly cultured in collagen I:
- Endothelial cells (including HUVEC)
- Fibroblasts
- Hepatocytes
- Chondrocytes
- Osteoclasts
- Osteoblasts
- Keratinocytes
- Corneal epithelium
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The key applications include the following:
- Collagen I induces microvascular endothelial cells to adopt a spindle-shaped morphology, in vitro, and to align into solid cord-like assemblies. Vascular endothelial cells can also form vessel-like, tubular structures when cultured on collagen I. Collagen I can be used for in vitro angiogenesis assays.
- Breast cancer stem cells can undergo differentiation when cultured on collagen I.
- Collagen I has been used for the culture of primary colon carcinoma cell lines; mouse liver progenitor cells have been cultured in 3D collagen I and rat pancreatic islets.
- Collagen I can act as a barrier in cell invasion assays, and has been used to study cell adhesion.
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Gibco Geltrex Matrix and collagen (rat tail and bovine) can be used as either a coating solution or a 3D gel matrix. CELLstart Substrate was developed to be used as a xeno-free coating matrix for only ESC applications (as a substitute for Gibco Geltrex Matrix or Matrigel Matrix, which are of animal origin). AlgiMatrix Matrix is a 3D scaffold-type matrix that does not support cell attachment but does provide a good environment for growing spheroids that can be easily harvested for downstream applications.
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