Collagenase A

Enzyme Commission (EC) Number:

3.4.24.3 ( BRENDA | IUBMB )

Кат. номер

10103586001

10103578001

11088793001

Collagenase is a protease which cleaves the triple-helical protein called collagen. Clostridium histolyticum produces six types of collagenases namely, , , , , and . These collagenases are active against connective tissue and show more reactivity towards gelatin than collagen. 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.
Collagenase degrades native collagen. Clostripain, trypsin-like enzymes, and neutral proteases also degrade other proteins.
Enzyme activity:
Collagenase activity: >0.15U/mg (according to Wunsch) (+25°C, 4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as the substrate)
Contaminating enzyme activities: trypsin, clostripain, and neutral proteolytic activity
Collagenase A has a balanced ratio of enzyme activities.
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) and for the preparation of single-cell suspensions for the establishment of primary cell culture systems. Collagenase A is recommended when yield, viability and functionality 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. However, suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically.
Unit Definition: Collagenase from Roche is assayed in Wnsch units (1 µmol of product formed per minute at +25 °C with Wnsch 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 Wnsch U/mg and 250 Mandl U/mg).
Contents
Lyophilizate, nonsterile
Activator: Ca2+
Inhibitors:
Collagenase inhibitors: EDTA, EGTA, Cys, His, DTT, 2-mercapto-ethanol
Collagenase is not inhibited by serum.
Clostripain inhibitors: TLCK
Trypsin inhibitors: aprotinin, trypsin inhibitor (egg white, soybean), serum
Working concentration: 0.5 to 2.5 mg/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.
Reconstitution in any balanced salt solution (e.g., HBSS)
For life science research only. Not for use in diagnostic procedures.
Shipping conditions may vary

Quality Level	100
sterility	non-sterile
form	lyophilized
packaging	pkg of 100 mg (10103578001), pkg of 2.5 g (11088793001), pkg of 500 mg (10103586001)
Торговая марка	Roche
optimum pH	6.0-8.0
shipped in	wet ice
storage temp.	2-8°C

RIDADR

NONH for all modes of transport