- 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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电泳缓冲液的另一个作用是使溶液具有一定的导电性,以利于DNA分子的迁移,例如,一般电泳缓冲液中应含有0.01-0.04mol/L的Na+离子,Na+离子的浓度太低时电泳速度变慢;太高时就会造成过大的电流使胶发热甚至熔化。
电泳缓冲液还有一个组分是EDTA,加入浓度为1-2mmol/L,目的是螯合Mg2+ 等离子,防止电泳时激活 DNA酶,此外还可防止Mg2+离子与核酸生成沉淀。
1mol/l Tirs-HCl(PH8.8) 0.6ml
10%SDS 2ml
50%甘油 5ml
2-巯基乙醇 0.5ml
1%溴酚兰 1ml
水 0.9ml
另外:SDS不好溶解建议加热溶解,如果要做非还原SDS-PAGE请将2-巯基乙醇 0.5ml换成超纯水即可
因为DNA和RNA链上的碱基都基本一致。而这两种上样缓冲液中含有的染料是二甲苯青和溴酚蓝,都可以与其碱基结合,跑出条带。
但是,如果使用DNA上样缓冲液,里面会夹杂一些RNA酶,这些酶有可能会影响到实验结果。使用前需要将RNA酶处理掉。
① 上样缓冲液中包括甘油,溴酚蓝,SDS等,甘油主要作用是防止样品漂浮出点样孔,溴酚蓝作为电泳指示剂,用来观察电泳进度,SDS用于一些核酸结合蛋白的变性解离.
②核酸染料与凝胶中核酸结合,以便于在紫外光下显示核酸条带.
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。
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