Storage: 2-8°C

UNSPSC Code: 12352200

RIDADR: NONH for all modes of transport

Features and Benefits:
Contents
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. However, suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically.

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

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

Physical form:
Lyophilizate, nonsterile.

Preparation Note:
Activator: Ca²⁺
Working concentration: 0.5 2.5mg/ml
Storage conditions (working solution): -15 to25°C
Roche recommends reconstituting only the amount of lyophilizate needed for immediate use. The reconstituted solution can be stored at15 to25°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 W×¼nsch) (+25°C, 4-phenyl-azobenzyl-oxycar<wbr>bonyl-Pro-Leu-Gly-Pro-D-A<wbr>rg 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).</wbr></wbr>

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.