COLLA-RO
Roche
Collagenase A
from Clostridium histolyticum
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About This Item
Recommended Products
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)
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.
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
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).
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.
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
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
Target Organs
Respiratory system
Storage Class Code
11 - Combustible Solids
WGK
WGK 1
Flash Point(F)
does not flash
Flash Point(C)
does not flash
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Articles
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.
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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