Collagenase D

from Clostridium histolyticum

Enzyme Commission number:

Quality Level






pkg of 100 mg (11088858001)
pkg of 2.5 g (11088882001)
pkg of 500 mg (11088866001)




(working concentration: 0.5 - 2.5 mg/ml)

optimum pH


shipped in

wet ice

storage temp.


General description

Collagenase D is prepared from Clostridium histolyticum cultures by filtration, ammonium sulfate precipitation, dialysis, and lyophilization.


Collagenase degrades native collagen. Clostripain, trypsin-like enzymes, and neutral proteases degrade other proteins as well.


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. However, suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically.

Features and Benefits

Lyophilizate, nonsterile

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.

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.


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 in any balanced salt solution (e.g., HBSS)

Other Notes

For life science research only. Not for use in diagnostic procedures.


NONH for all modes of transport

Charles J Benck et al.
Journal of visualized experiments : JoVE, (107)(107), e53638-e53638 (2016-02-11)
Dissociating murine skin into a single cell suspension is essential for downstream cellular analysis such as the characterization of infiltrating immune cells in rodent models of skin inflammation. Here, we describe a protocol for the digestion of mouse skin in...
Chibuzor Uchea et al.
Toxicology research, 4(2), 494-507 (2015-04-22)
Prediction of xenobiotic fate in fish is important for the regulatory assessment of chemicals under current legislation. Trout hepatocyte spheroids are a promising
Adrienn Angyal et al.
International immunology, 22(2), 71-80 (2009-12-03)
Fcgamma receptors (FcgammaRs) play an essential role in the regulation of immune response due to their ability to bind immune complexes. Activating FcgammaRs may facilitate antigen presentation and dendritic-cell maturation, while in the late phase of the immune response, the...
Yu Fan et al.
Mediators of inflammation, 2014, 397815-397815 (2014-05-09)
Clinical studies suggested thatandrogen might be associated with infiltrating T cells in prostate of benign prostatic hyperplasia (BPH) patients, but detail of T-cell subset and mechanism still remained unclear. The present study tested the hypothesis that intraprostatic 5 α -dihydrotestosterone...
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.
Read More
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.
Read More

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service

Social Media

LinkedIn icon
Twitter icon
Facebook Icon
Instagram Icon


Research. Development. Production.

We are a leading supplier to the global Life Science industry with solutions and services for research, biotechnology development and production, and pharmaceutical drug therapy development and production.

© 2021 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved.

Reproduction of any materials from the site is strictly forbidden without permission.