产品名称

Collagenase from Clostridium histolyticum

别 称

Clostridiopeptidase A

含 量

Type IV, 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

浅米色至米褐色和淡棕色至深棕色的粉末

溶解性(25°C)

TESCA buffer (50mM TES, 0.36mM Calcium chloride, pH7.4): soluble 0.05-0.1mg/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. Effective release of cells from tissue requires the action of 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. The pH optimum is 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 are used for digestion. The duration of reaction  varies from 15 minutes to several hours and yields satisfactory cell dissociation without causing too much cell death. Krebs Ringer buffer with calcium and BSA is preferred and Zn₂₊ is required for 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.This enzyme is tested for suitability for the release of hepatocytes, at approx. 1mg/mL conc, in a total volume of 100mL for each rat liver.

样品准备说明

Solutions are typically prepared at 1-2mg/mL in TESCA buffer (containing 50mM TES, 0.36mM Calcium chloride, pH7.4 at 37°C. This product also contains clostripain, neutral protease and tryptic  activities.

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 5hours 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 1.0 μmole 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