Sigma-Aldrich Collagenase B 500mg

Biological Source

Clostridium histolyticum

Quality Level

100

Sterility

non-sterile

Form

lyophilized

Packaging

pkg of 100 mg (11088807001)
pkg of 2.5 g (11088831001)
pkg of 500 mg (11088815001)

Manufacturer/Tradename

Roche

Concentration

0.5-2.5 mg/mL

Optimum pH

6.0-8.0

Application(S)

sample preparation

Storage Temp.

2-8°C

Related Categories

Collagenase

Roche® Life Science Products

Tissue Dissociation

General Description

Collagenase B is prepared fromClostridium histolyticumcultures by filtration, ammonium sulfate precipitation, dialysis, and lyophilization. Collagenase fromClostridium histolyticumis a collagenolytic enzyme. It interacts with collagen-like substances and is responsible for the degradation of collagen into small peptides.It 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 B is recommended when yield and viability are important.

Specificity

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

Application

Clostridium collagenasefrom 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.
Collagenase B has been used for preparation of cells from mouse aortas and colorectal cancer tissue.

Specifications

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 total proteolytic activity
Collagenase B has a normal to high collagenase activity and a higher than average clostripain activity (usually >10U/mg).

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), serum

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).

Preparation Note

Activator:Ca²⁺
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

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

Other Notes

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

Pictograms

GHS07,GHS08

Signalword

Danger

Hcodes

H315 - H319 - H334 - H335

Pcodes

P261 - P264 - P280 - P284 - P304 + P340 + P312 - P342 + P311

Hazard Classifications

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

Target_Organs

Respiratory system

Storage Class

11 - Combustible Solids

Wgk_Germany

WGK 1

Flash_Point_F

does not flash

Flash_Point_C

does not flash