C1889
Collagenase from Clostridium histolyticum
0.2 μm filtered, suitable for release of physiologically active rat hepatocytes, Type IV-S, 0.5-5.0 FALGPA units/mg solid, ≥125 CDU/mg solid
Synonym(s):
Clostridiopeptidase A
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| Pack Size | SKU | Availability | Price |
|---|---|---|---|
| 50 mg | Check Cart for Availability | $116.00 |
About This Item
CAS Number:
UNSPSC Code:
12352204
NACRES:
NA.54
EC Number:
232-582-9
MDL number:
Specific activity:
≥125 CDU/mg solid, 0.5-5.0 FALGPA units/mg solid
Quality Level
sterility
0.2 μm filtered
type
Type IV-S
form
powder
specific activity
≥125 CDU/mg solid, 0.5-5.0 FALGPA units/mg solid
mol wt
68-130 kDa
suitability
suitable for release of physiologically active rat hepatocytes
application(s)
diagnostic assay manufacturing
storage temp.
−20°C
General description
Clostridium histolyticum consists of two classes of collagenases, which cleaves at multiple cleavage sites within the collagen triple helix. It is more effective than mammalian collagenases. Clostridial collagenase is implicated in bacterial invasion in gas gangrene.
Application
Collagenase has been used in the preparation of arterial tissue for the study of Advanced Glycosylation End Products (AGE). The enzyme has also been used along with other proteases for the disaggregation of human tumor, mouse kidney, human brain, lung epithelium and many other tissues. It is also effective in liver and kidney perfusion studies, digestion of pancreas, and isolation of nonparenchymal hepatocytes. When this enzyme is tested for suitability for the release of hepatocytes. The collagenase is used at approximately 1 mg/mL in a total volume of 100 mL for each rat liver.
Collagenase from Clostridium histolyticum has been used to dissociate tissue samples such as spleens[1] and testicular tissue.
Biochem/physiol Actions
Collagenase is activated by four gram atom calcium per mole enzyme. It is inhibited by ethylene glycol-bis(beta-aminoethyl ether) - N, N, N′,N′-tetraacetic acid, beta-mercaptoethanol, glutathione, thioglycolic acid and 8-hydroxyquinoline.
Effective release of cells from tissue requires the action of both collagenase enzymes and the neutral protease. Collagenase is activated by four gram atom calcium (Ca2+) per mole enzyme. The culture filtrate is thought to contain at least 7 different proteases ranging in molecular weight from 68-130 kDa (pH optimum: 6.3-8.8). The enzyme is typically used to digest the connective components in tissue samples to liberate individual cells. Collagenase treatment can cause some cells to die. Typically, concentrations varying from 0.1 to 5 mg/mL is used for digestion. The duration of reaction varies from 15 minutes to several hours, which enables efficient cell dissociation without causing too much cell death. Krebs Ringer buffer with calcium and BSA is preffered and Zn2+ is required for its activity.
Preparation Note
Prepared from Type IV (C5138). A stock solution may be prepared by dissolving 0.05-0.1 mg/mL of collagenase in 50 mM TES buffer containing 0.36 mM calcium chloride (TESCA) (pH 7.4) at 37 °C. This product also contains clostripain, nonspecific neutral protease, and tryptic activities.
Other Notes
One collagen digestion unit (CDU) liberates peptides from collagen from bovine achilles tendon equivalent in ninhydrin color to 1.0 μmole of leucine in 5 hours at pH 7.4 at 37 °C in the presence of calcium ions. One FALGPA hydrolysis unit hydrolyzes 1.0 μmole of furylacryloyl-Leu-Gly-Pro-Ala per min at 25°C. One Neutral Protease unit hydrolyzes casein to produce color equivalent to 1.0 μmole of tyrosine per 5 hr at pH 7.5 at 37°C. One Clostripain Unit hydrolyzes 1.0 μmole of BAEE per min at pH 7.6 at 25°C in the presence of DTT.
Disclaimer
As supplied, this product is stable for one year at -20°C. There is no loss in FALGPA or protease activity in 30 days at 37°C, 50°C and -20°C. Solutions of crude collagenase are stable if frozen quickly in aliquots (at 10 mg/mL) and kept frozen at -20°C. Further freeze-thaw cycles will damage the solution. The product retains 100% activity over 7 hours when held on ice.
1 of 1
This Item | |||
|---|---|---|---|
| specific activity ≥125 CDU/mg solid, 0.5-5.0 FALGPA units/mg solid | specific activity ≥125 CDU/mg solid, 0.5-5.0 FALGPA units/mg solid | specific activity ≥125 CDU/mg solid, 0.5-5.0 FALGPA units/mg solid | specific activity ≥125 CDU/mg solid, 0.5-5.0 FALGPA units/mg solid |
| suitability suitable for release of physiologically active rat hepatocytes | suitability - | suitability - | suitability - |
| form powder | form powder | form powder | form essentially salt-free, lyophilized powder |
| application(s) diagnostic assay manufacturing | application(s) diagnostic assay manufacturing | application(s) diagnostic assay manufacturing | application(s) diagnostic assay manufacturing |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
| sterility 0.2 μm filtered | sterility - | sterility - | sterility - |
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signalword
Danger
hcodes
Hazard Classifications
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
target_organs
Respiratory system
Storage Class
11 - Combustible Solids
wgk
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
dust mask type N95 (US), Eyeshields, Faceshields, Gloves
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Protocols
To measure collagenase activity, N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala is used in a continuous spectrophotometric rate determination at 345 nm. Collagenase hydrolyzes collagen peptide bonds.
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Global Trade Item Number
| SKU | GTIN |
|---|---|
| C1889-50MG | 04061833468272 |