产品名称

Collagenase from Clostridium histolyticum

别 称

Clostridiopeptidase A

含 量

Type IV-S, 0.5-5.0 FALGPA units/mg solid, ≥125 CDU/mg solid

CAS  NO.

9001-12-1

MDL  number

MFCD00130830

适用范围

生物试剂

l  理化信息

外 观

粉末

l  生物学信息

生物活性/药理作用

Collagenase is activated by four gram atom calcium per mole enzyme. It is inhibited by ethylene glycol-bis(beta-aminoethyl ether) - N, N, N′,N′-tetraacetic acid, beta-mercaptoethanol, glutathione, thioglycolic acid and 8-hydroxyquinoline. Effective release of cells from tissue requires the action of both collagenase enzymes and the neutral protease. Collagenase is activated by four gram atom calcium (Ca₂₊⁾ per mole enzyme. The culture filtrate is thought to contain at least 7 different proteases ranging in molecular weight from 68-130 kDa (pH optimum: 6.3-8.8). The enzyme is typically used to digest the connective components in tissue samples to liberate individual cells. Collagenase treatment can cause some cells to die. Typically, concentrations varying from 0.1 to 5mg/mL is used for digestion. The duration of reaction varies from 15 minutes to several hours, which enables efficient cell dissociation without causing too much cell death. Krebs Ringer buffer with calcium and BSA is preffered and Zn₂₊ is required for its activity.

应 用

Collagenase has been used in the  preparation of arterial tissue for the study of Advanced Glycosylation End Products (AGE). The enzyme has also been used along with other proteases for the disaggregation of human tumor, mouse kidney, human brain, lung epithelium and many other tissues. It is also effective in liver and kidney perfusion studies, digestion of pancreas, and isolation of nonparenchymal hepatocytes. When this enzyme is tested for suitability for the release of hepatocytes. The collagenase is used at approximately 1mg/mL in a total volume of 100mL for each rat liver.

注 意

As supplied, this product is stable for one year at -20°C. There is no loss in FALGPA or protease activity in 30 days at 37°C, 50°C and -20°C. Solutions of crude collagenase are stable if frozen quickly in aliquots (at 10mg/mL) and kept frozen at -20°C. Further freeze-thaw cycles will damage the solution.The product retains 100% activity over 7 hours when held on ice.

有效单位定义

One collagen digestion unit (CDU) liberates peptides from collagen from bovine achilles tendon equivalent in ninhydrin color to 1.0μmole of leucine in 5 hours at pH 7.4 at 37 °C in the presence of calcium ions. One FALGPA hydrolysis unit hydrolyzes 1.0 μmole of furylacryloyl-Leu-Gly-Pro-Ala per min at 25°C. One Neutral Protease unit hydrolyzes casein to produce color equivalent to 1.0μmole of tyrosine per 5 hr at pH7.5 at 37°C. One Clostripain Unit hydrolyzes 1.0μmole of BAEE per min at pH7.6 at 25°C in the presence of DTT.

l  存储

存储温度

-20°C

l  注意事项及免责声明

本产品仅用于实验研究，不得作为药物使用，不得用于家用或其它用途。

l  参考文献

1.    http://www.drugbank.ca

2.    https://ncit.nci.nih.gov

3.    https://www.ncbi.nlm.nih.gov