- Overview
- Data/Specifications
- Literature/Support
- How To Use
- Related Products
Overview
Type I collagen is the most abundant collagen and is found in connective tissues including tendon, ligament, dermis and blood vessel. It is the major component and the primary determinant of tensile strength of the extracelluar matrix (ECM). It is widely used as a thin layer on tissue-culture surfaces to enhance the attachment and proliferation of a variety of cells including endothelial cells, fibroblasts, hepatocytes, epithelial cells etc. In addition, collagen I can self-assemble into a 3-D superamolecular gel in vitro, making it an ideal biological scaffold to promote more in vivo-like cellular morphology and function.
Symbols/Related Terms:
- COL1A1
- COL1A2
- osteogenesis imperfecta
Data/Specifications
Description: Purified Rat Collagen Type I (Atelocollagen) from rat tail tendon
Form: Lyophilized, salt free, 30 mg/ vial
Sterility: Passed sterility test for bacteria and fungi
Source: Rat tail tendons
Purity: Rat collagen Type I > 90% / Rat collagen type III <10%
Purification: Collagen was extracted from dissected tissue into diluted acetic acid after mild pepsin treatment. Collagen type I was purified by using differential salt precipitation.
Reconstitution: Dissolve in 20 mM acetic acid. Suspension should be shaken or stirred for several hours or overnight between 2-8 °C. Material is dissolved when it appears homogeneous, with no schlieren pattern (light refraction due to differing protein concentrations in the suspensions). Recommended concentration 1-5 mg/mL.
Storage: Collagen dissolved in acetic acid is stable at 4°C for 1 month. Lyophilized collagen long term storage at -20°C or lower.
Applications: Cell culture (coating or 3D gel)
Literature/Support
Purified Rat Collagen Type I (Atelocollagen) from rat tail tendon Insert (PDF)
Note: inserts are for review only. Please use the insert that was sent with the product to ensure that the correct revision insert is being used for the product purchased.
How To Use
Dissolve in 20 mM acetic acid. Suspension should be shaken or stirred for several hours or overnight between 2-8°C. Material is dissolved when it appears homogeneous, with no schlieren pattern (light refraction due to differing protein concentrations in the suspensions). Recommended concentration 1-5 mg/mL.
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蛋白质上样缓冲液中的SDS和DTT分别可以屏蔽蛋白质电荷以及破坏二硫键，避免形成多聚体。
DNA上样缓冲液主要是起沉降和指示作用，不可用于蛋白，否则会影响蛋白相对分子量定量。
同样蛋白上样缓冲液中的tris-HCl浓度过高也会使溴酚蓝条带扭曲。
2乘SDS-PAGE上样缓冲液（20ml体系）
1.0mol/L Tris-HCL (pH 6.8) 2ml
1.0mol/L DTT 4ml
SDS 0.8g
溴芬蓝 0.04g
甘油 4ml
dd水 定容至20ml
4℃冰箱保存，可以不分装，我们实验室一般用三个月以上
先配1.0mol/L的DTT,双蒸无菌水溶解4℃冰箱保存,配上样缓冲液的时候加入就可以了,都混匀保存也没问题的
1。称量氨基丁三醇 242g，Na2EDTA.2H2O 37.2g 于1L烧杯中；
2。向烧杯中加入约600ml去离子水，充分搅拌均匀；
3。加入57.1ml的冰乙酸，充分溶解；
4。用NaOH调pH至8.3，加去离子水定容至1L后，室温保存。
使用时稀释50倍 即1×TAE Buffer
10×TBE Buffer配制方法：
1。称量氨基丁三醇 108g，Na2EDTA.2H2O 7.44g，硼酸55g 于1L烧杯中；
2。加入约700ml去离子水，搅拌均匀；
3。用NaOH调pH至8.3，加去离子水定容至1L后，室温保存。
使用时稀释10倍 即1×TBE Buffer
0.5 M tris-HCl (pH 6.8) 20％
丙三醇 20％
20％SDS 20％
0.1％溴酚兰 5％
2-巯基乙醇 10％
双蒸水 25％
不调PH值，PH在配0.5 M tris-HCl 时已调，缓冲液呈蓝紫色。
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