This product is not specifically tested for use in 'cell culture', however it is tested for cell dissociation. The testing falls into the broad scope of cell culture in that the product is not cytotoxic and is enzymatically capable of preparing a single cell suspension. Details may be review in the PROPERTIES section as well as the Certificate of Analysis. See the link below to review a sample document:
https://www.sigmaaldrich.com/certificates/sapfs/PROD/sap/certificate_pdfs/COFA/Q14/C2674-100MG0000227617.pdf
C2674
Collagenase from Clostridium histolyticum
lyophilized powder, ≥125 CDU/mg solid (CDU = collagen digestion units), 0.5-5.0 FALGPA units/mg solid
Synonym(s):
Clostridiopeptidase A
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100 MG
$159.00
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$439.00
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$747.00
About This Item
CAS Number:
UNSPSC Code:
12352204
NACRES:
NA.75
EC Number:
232-582-9
MDL number:
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Product Name
Collagenase from Clostridium histolyticum, lyophilized powder, ≥125 CDU/mg solid (CDU = collagen digestion units), 0.5-5.0 FALGPA units/mg solid
biological source
Clostridium histolyticum
type
Type I-A
form
lyophilized powder
specific activity
≥125 CDU/mg solid (CDU = collagen digestion units)
0.5-5.0 FALGPA units/mg solid
mol wt
68-130 kDa
concentration
1 mg/mL
technique(s)
cell culture | mammalian: suitable
single cell analysis: suitable
pH
6.3-8.8
shipped in
ambient
storage temp.
−20°C
Quality Level
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1 of 4
This Item | C0255 | C9263 | C2139 |
|---|---|---|---|
| specific activity ≥125 CDU/mg solid (CDU = collagen digestion units), 0.5-5.0 FALGPA units/mg solid | specific activity ≥500 CDU/mg solid (CDU = collagen digestion units) | specific activity ≥1 FALGPA units/mg solid, >125 CDU/mg solid | specific activity ≥125 CDU/mg solid, 0.5-5.0 FALGPA units/mg solid |
| technique(s) cell culture | mammalian: suitable, single cell analysis: suitable | technique(s) - | technique(s) - | technique(s) - |
| biological source Clostridium histolyticum | biological source - | biological source - | biological source Clostridium histolyticum |
| form lyophilized powder | form lyophilized powder | form powder | form powder |
| concentration 1 mg/mL | concentration - | concentration - | concentration - |
| shipped in ambient | shipped in - | shipped in - | shipped in - |
Application
Collagenase from Clostridium histolyticum is used for the following applications:
This product is suitable for the disaggregation of human tumor, mouse kidney, human adult and fetal brain, lung and many other epithelia tissues. It has also been shown to be effective in liver and kidney perfusion studies, digestion of pancreas, isolation of nonparenchymal rat liver cells and hepatocyte preparation. Collagenase has also been used in the preparation of arterial tissue for the study of Advanced Glycosylation End Products. This enzyme has been tested for the release of heptatocytes at a concentration of approximately 1mg/mL. Concentrations for digestion range from 0.1 to 5mg/mL.
Suitable for use in preparation of single cell suspension for sequencing.
- Sertoli cell isolation[1]
- Used in the comparison of enzymatic methods[2]
- Used during tissue preparations for immunocytochemistry[3]
- Testicular sperm extraction[4]
- Preparation of single cell suspensions[5]
- Immunofluorescence[6]
This product is suitable for the disaggregation of human tumor, mouse kidney, human adult and fetal brain, lung and many other epithelia tissues. It has also been shown to be effective in liver and kidney perfusion studies, digestion of pancreas, isolation of nonparenchymal rat liver cells and hepatocyte preparation. Collagenase has also been used in the preparation of arterial tissue for the study of Advanced Glycosylation End Products. This enzyme has been tested for the release of heptatocytes at a concentration of approximately 1mg/mL. Concentrations for digestion range from 0.1 to 5mg/mL.
Suitable for use in preparation of single cell suspension for sequencing.
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.
The collagenase product is a mixture of enzymes secreted by C. histolyticum, with different products differentiated by the relative ratios of the 10-18 components found in the secreted enzymes. The main components are two collagenases, clostripain, and a neutral protease. The synergistic action of these enzymes degrade collagen and other intracellular materialThe action of both collagenase enzymes and the neutral protease is necessary for effective release of cells from tissue. Various types of collagen are the natural substrates for collagenase.
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.
General description
Collagenase is a protease which cleaves the triple-helical protein called collagen. There are three types of tissue collagenases, and these belong to the matrix metalloproteinases (MMP) family.[7] 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.[8] Crude collagenase mixtures contain two major enzyme types namely, collagenase and clostripain.[9]
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.
Preparation Note
This collagenase is obtained from the culture filtrate of Clostridium histolyticum. The culture filtrate is thought to contain at least 7 different proteases ranging in molecular weight from 68-130 kDa. This product is Type I-A. Solutions are typically prepared at 1-2 mg/mL in TESCA buffer (containing 50 mM TES, 0.36 mM Calcium chloride, pH 7.4 at 37°C.
used together
Product No.
Description
Pricing
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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Treatment of rat liver plasma membranes with various commercial preparations of crude collagenase from Clostridium histolyticum at concentrations as low as 1 mug/ml, resulted in activation of the adenylate cyclase system. Maximal activation occurred at 50 to 100 mug/ml of
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Rift Valley fever virus (RVFV) causes severe disease in livestock concurrent with zoonotic transmission to humans. A subset of people infected with RVFV develop encephalitis, and significant gaps remain in our knowledge of how RVFV causes pathology in the brain.
Articles
Quantitative and qualitative western blotting to validate knockdown by esiRNA.
Discover pre-mixed collagenase enzyme blends with DNase I, Dispase II, Elastase, and Hyaluronidase and gently dissociate animal tissues in vitro.
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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