产品名称

Collagenase from Clostridium histolyticum

别 称

Clostridiopeptidase A

含 量

Type XI, 2-5 FALGPA units/mg solid, ≥800 CDU/mg solid

CAS  NO.

9001-12-1

MDL  number

MFCD00130830

适用范围

生物试剂,  Suitable for isolation of rat pancreatic islet cells.

l  理化信息

外 观

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 the reaction varies from 15 minutes to several hours to yield a satisfactory efficiency of cell dissociation without causing too much cell death. Krebs Ringer buffer with calcium and BSA is preffered and Zn₂₊ is required for activity. This enzyme is tested for the release of hepatocytes at approximately 1mg/mL concentration, in a total volume of 100mL for each rat liver.

应 用

Clostridiopeptidase A has been used to study the avoidance of the progression of necrotic tissue in neonates as well as to evaluate the effect of clostridiopeptidase A collagenase (CAC) in a rat model of postoperative peritoneal adhesion. Clostridiopeptidase A has also been used in a study to describe a quantitative method to assess small amounts of collagen d on MALDI-TOF(matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry (MS) subsequent to digestion of collagen. This product has been used in the isolation of pancreatic ductal adenocarcinoma (PDAC) cell lines and the tissue culture of pancreatic fibroblasts.

样品准备说明

A stock solution may be prepared by dissolving 0.05-0.1mg/mL of collagenase in 50mM TES buffer containing 0.36mM calcium chloride (TESCA), pH7.4, at 37°C.

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 pH 7.6 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