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COLLA-RO

Roche

Collagenase A

from Clostridium histolyticum

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Synonym(s):
Collagenase
Enzyme Commission number:

biological source

Clostridium histolyticum

Quality Level

sterility

non-sterile

form

lyophilized

specific activity

>0.15 U/mg lyophilizate (Collagenase activity)

packaging

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

manufacturer/tradename

Roche

concentration

1 mg/mL (0.1%, w/v)

color

brown

optimum pH

6.0-8.0

solubility

water: soluble

suitability

suitable for PCR

NCBI accession no.

UniProt accession no.

application(s)

cell analysis
life science and biopharma
sample preparation

foreign activity

Clostripain 9.489 U/mg
Proteases 6.067 U/mg (Azocoll )
Trypsin 0.144 U/mg (with BAEE)

storage temp.

2-8°C

Gene Information

Clostridium histolyticum ... COLA_1(1668506084)

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This Item
COLLDISP-ROCOLLH-ROCOLLD-RO
vibrant-m

COLLA-RO

Collagenase A

vibrant-m

COLLDISP-RO

Collagenase/Dispase®

vibrant-m

COLLH-RO

Collagenase H

vibrant-m

COLLD-RO

Collagenase D

specific activity

>0.15 U/mg lyophilizate (Collagenase activity)

specific activity

>0.8 U/mg lyophilizate, pH 7.5, 37 °C (casein as substrate, Dispase)

specific activity

-

specific activity

-

biological source

Clostridium histolyticum

biological source

enzyme from Bacillus polymyxa ( Dispase ), enzyme from bacterial ( Collagenase from Vibrio alginolyticus)

biological source

Clostridium histolyticum

biological source

Clostridium histolyticum

application(s)

cell analysis
life science and biopharma
sample preparation

application(s)

life science and biopharma
sample preparation

application(s)

life science and biopharma

application(s)

cell analysis

concentration

1 mg/mL (0.1%, w/v)

concentration

1 mg/mL (Collagenase: 0.1U/mL, Dispase: 0.8U/mL)

concentration

~1 mg/mL (Isolation of rat hepatocytes), ~2 mg/mL (Isolation of adipocytes)

concentration

(working concentration: 0.5 - 2.5 mg/ml)

form

lyophilized

form

lyophilized

form

lyophilized

form

lyophilized

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.

Specificity

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.

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, umbilical cord)
  • preparation of single-cell suspensions for the establishment of primary cell culture systems. Collagenase A is recommended when yield, viability, and functionality are important.
  • preparation of cells from many types of tissue, such as hepatocytes, adipocytes, pancreatic islets, epithelial cells, muscle cells, endothelial cells, etc. However, the suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically.

Unit Definition

Contents
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).

Preparation Note

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

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

Storage and Stability

Keep container tightly closed in a dry and well-ventilated place

Other Notes

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

pictograms

Exclamation markHealth hazard

signalword

Danger

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


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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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