Collagenase A

from Clostridium histolyticum

Enzyme Commission number:

Quality Level






pkg of 100 mg (10103578001)
pkg of 2.5 g (11088793001)
pkg of 500 mg (10103586001)



optimum pH


shipped in

wet ice

storage temp.


General description

Collagenase is a protease which cleaves the triple-helical protein called collagen. 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. 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 Wünsch) (+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

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).
Lyophilizate, nonsterile

Preparation Note

Activator: Ca2+
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)

Other Notes

For life science research only. Not for use in diagnostic procedures.
Shipping conditions may vary


Exclamation markHealth hazard

Signal Word


Hazard Statements


Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

Target Organs

Respiratory system


11 - Combustible Solids

Flash Point(F)

does not flash

Flash Point(C)

does not flash

Certificate of Analysis

Certificate of Origin

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Enzyme Explorer Key Resource: Collagenase Guide.Collagenases, enzymes that break down the native collagen that holds animal tissues together, are made by a variety of microorganisms and by many different animal cells.
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Related Content
Collagenase Guide.Collagenases, enzymes that break down the native collagen that holds animal tissues together, are made by a variety of microorganisms and by many different animal cells.
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