Collagenase A

Description

• General description: Collagenase is a protease which cleaves the triple-helical protein called collagen.[1] Clostridium histolyticum produces six types of collagenases namely, α, β, γ, δ, ε and λ. These collagenases are active against connective tissue and show more reactivity towards gelatin than collagen.[2] 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.[3]
• Specificity: Collagenase degrades native collagen. Clostripain, trypsin-like enzymes, and neutral proteases also degrade other proteins.Enzyme activity:Collagenase activity: >0.15U/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 neutral proteolytic activityCollagenase A has a balanced ratio of enzyme activities.
• 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, umbilical cord) preparation of single-cell suspensions for the establishment of primary cell culture systems.[4] Collagenase A is recommended when yield, viability, and functionality are important.[5][6]preparation of cells from many types of tissue, such as hepatocytes, adipocytes, pancreatic islets, epithelial cells, muscle cells, endothelial cells, etc. However, the suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically.[7][8]
• Unit Definition: ContentsLyophilizate, nonsterile
• Preparation Note: Activator: Ca2+Inhibitors:Collagenase inhibitors: EDTA, EGTA, Cys, His, DTT, 2-mercapto-ethanolCollagenase is not inhibited by serum.Clostripain inhibitors: TLCKTrypsin inhibitors: aprotinin, trypsin inhibitor (egg white, soybean), serumWorking concentration: 0.5 to 2.5 mg/mlStorage conditions (working solution): -15 to -25 °CRoche 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.
• Reconstitution: Reconstitution in any balanced salt solution (e.g., HBSS)
• Storage and Stability: Keep container tightly closed in a dry and well-ventilated place
• Other Notes: For life science research only. Not for use in diagnostic procedures.