l 基本信息 |
产品名称 | Collagenase from Clostridium histolyticum |
别 称 | Clostridiopeptidase A |
含 量 | Type IA, 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 | 浅米色至米褐色和浅棕色至黑色冻干粉末（基本无盐） |
溶解性(25°C) | TESCA buffer (50mM TES, 0.36mM Calcium chloride, pH7.4): soluble |
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. The pH optimum ranges from 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 for 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. The enzyme recognizes the sequence -R-Pro-8-X-Gly-Pro-R-, where X is most often a neutral amino acid. Ethylene glycol-bis(β-aminoethyl
ether)-N,N,N′,N′-tetraacetic acid (EGTA)4; β-mercaptoethanol; glutathione (reduced); thioglycolic acid, sodium; 2,2′-dipyridyl; 8-hydroxyquinoline, cysteine are known to inhibit the enzyme activity. |
应 用 | The enzyme has also been used along with other proteases for the disaggregation of human tumor, mouse kidney, human brain, and lung epithelial tissues. It is also useful in liver and kidney perfusion studies, digestion of pancreas, isolation of nonparenchymal rat liver cells and hepatocytes. The enzyme from Sigma has been used for the digestion of type I collagen from bovine trabecular and cortical bones.It has also been used as a positive control in FALGPA assay on Group B Streptococci
cells. |
样品准备说明 | This product is 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 1.0μmole 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 |
