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Roche 11088882001 Collagenase D胶原酶D
公司简介
罗氏总部位于瑞士巴塞尔，在制药和诊断领域是世界领先的以研发为基础，以创新为驱动的健康医疗公司之一。作为全球最大的生物技术公司之一，罗氏在抗肿瘤、抗病毒、炎症、代谢和中枢神经系统领域拥有切实有效的差异化药品；同时罗氏也是体外诊断和基于组织的肿瘤诊断的市场领导者，以及糖尿病管理领域的先驱者。罗氏倡导的“个性化医疗”方案旨在提供创新的药品和诊断工具，从而切实改善人类的健康水准、生活质量以及患者的生存期。
在成功收购德国宝灵曼公司（Boehringer Mannheim GmbH.）后，成立了专业服务于科学研究领域用户的罗氏诊断生命科学部（Roche Life Science），凭借半个多世纪在生命科学研究领域积累的经验，罗氏诊断生命科学部已成为在此领域中具重要地位的试剂及系统供应商之一。罗氏诊断生命科学部还为生命科学行业、生物制药行业和诊断行业提供所需的原料、试剂组分和试剂盒，罗氏诊断生命科学部为客户进行量身定制产品，满足客户个性化需求。
产品介绍
Features and Benefits
Contents
Lyophilizate, nonsterile
General description
Collagenase D is prepared from Clostridium histolyticum cultures by filtration, ammonium sulfate precipitation, dialysis, and lyophilization.
Other Notes
仅用于生命科学研究。不可用于诊断。
Preparation Note
Activator: Ca2+
Working concentration: 0.5 - 2.5mg/ml
Storage conditions (working solution): -15 to -25°C
Roche recommends reconstituting only the amount of lyophilizate needed for immediate use. The reconstituted solution can be stored at -15 to -25°C for up to one week. Avoid repeated freezing and thawing since activity decreases after reconstitution.
Inhibitors:
Collagenase inhibitors: EDTA, EGTA, Cys, His, DTT, 2-mercaptoethanol
Collagenase is not inhibited by serum.
Clostripain inhibitors: TLCK
Trypsin inhibitors: aprotinin, trypsin inhibitor (egg white, soybean)
Enzyme activity:
Collagenase activity: >0.15 U/mg (according to Wünsch) (+25°C, 4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as the substrate)
Contaminating enzyme activities: trypsin, clostripain, and total proteolytic activity
Collagenase D has a normal to high collagenase activity and very low tryptic activity (usually <0.2 units/mg, BAEE as substrate).
Reconstitution
Reconstitution in any balanced salt solution (e.g., HBSS)
Specificity
Collagenase degrades native collagen. Clostripain, trypsin-like enzymes, and neutral proteases degrade other proteins as well.
Unit Definition
Collagenase from Roche is assayed in Wünsch units (1 μmol of product formed per minute at +25 °C with Wünsch substrate).
Frequently, collagenase activities are given in Mandl units (1 μmol leucine liberated from collagen in 5 hours at +37 °C).
Unfortunately, there is no consistent conversion factor between the two units of activity, since the Mandl unit depends, in part, on the concentration of contaminating proteases in the collagenase preparation, an indefinable variable. A purer collagenase preparation would actually give a lower specific activity in Mandl units than a crude preparation. Clostridium preparations typically give conversion factors of approximately 1:1800 (e.g., a particular lot of Clostridium collagenase contained approximately 0.15 Wünsch U/mg and 250 Mandl U/mg).
Physical form
Lyophilizate, nonsterile.
Application
Collagenase from C. histolyticum is widely used for the disaggregation of many types of tissues (e.g., lung, heart, muscle, bone, adipose tissue, liver, kidney, cartilage, mammary gland, placentae, blood vessels, brain, tumors) and for the preparation of single cell suspensions for the establishment of primary cell culture systems. Collagenase D is recommended when functionality and integrity of cell-surface proteins are important.
Clostridium collagenase from Roche has been used to prepare cells from many types of tissue, such as hepatocytes, adipocytes, pancreatic islets, epithelial cells, muscle cells, endothelial cells, etc.[4][3][1][2] However, suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically.