产品名称

Collagenase from Clostridium histolyticum

一般描述

Collagenase is a protease which cleaves the  triple-helical protein called collagen. There are three types of tissue collagenases, and these belong to the matrix lloproteinases (MMP) family. Collagenase obtained from Clostridium histolyticum has a very strong activity, as it digests collagen from both ends, at temperatures as low as 4-10°C. Crude collagenase mixtures contain two major enzyme types namely, collagenase and clostripain.

别 称

Clostridiopeptidase A

含 量

Type I, 0.25-1.0 FALGPA units/mg solid,  ≥125 CDU/mg solid

CAS  NO.

9001-12-1

分子量

mol wt 68,000 to 125,000

MDL  number

MFCD00130830

适用范围

生物试剂

l  理化信息

物理形态

基本上无盐的冻干粉末

溶解性(25°C)

TESCA buffer (50mM TES, 0.36mM Calcium chloride, pH7.4): soluble 1-2 mg/mL at 37°C

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.  The collagenase product is a mixture of enzymes secreted by C. histolyticum, with different products differentiated by the relative ratios of the 10-18 components found in the secreted enzymes. The main components are two collagenases, clostripain, and a neutral protease. The synergistic action of these enzymes  degrade collagen and other intracellular material The action of both collagenase enzymes and the neutral protease is necessary for effective release of cells from tissue. Various types of collagen are the natural substrates for collagenase.

应 用

Collagenase from Clostridium histolyticum has extensive use in biological studies, where it is used to prepare isolated cell suspensions.This product is suitable for the disaggregation of human tumor, mouse kidney, human adult and fetal brain, lung and many other epithelia tissues. It has also been shown to be effective in liver and kidney perfusion studies, digestion of pancreas, isolation of nonparenchymal rat liver cells and hepatocyte preparation. Collagenase has also been used in the preparation of arterial tissue for the study of Advanced Glycosylation End  Products. This enzyme has been tested for the release of heptatocytes at a concentration of approximately 1mg/mL. Concentrations for digestion range  from 0.1 to 5mg/mL.

样品准备说明

This product is an equivalent to first 40% ammonium sulfate fraction of Mandl, I., et al., J. Clin. Invest., 32, 1323 (1953). Solutions are typically prepared at 1-2mg/mL in TESCA buffer (containing 50mM TES, 0.36mM Calcium chloride, pH7.4 at 37°C.

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 pH7.4 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