Collagenase would be soluble in PBS, however it is recommended to dissolve the enzyme in TESCA buffer containing 50 mM TES, 0.36 mM Calcium chloride. Aliquotes may be stored up to 1 year at -20°C. Avoid repeated freeze thaw cycles. Please see the link below to the product information sheet for additional details.
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/321/500/c6885pis.pdf
C6885
Collagenase from Clostridium histolyticum
suitable for release of physiologically active rat epididymal adipocytes, Type II, 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 |
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| 25 mg | Available to ship TODAYfromMILWAUKEE | $70.10 | |
| 100 mg | Available to ship TODAYfromMILWAUKEE | $116.00 | |
| 500 mg | Available to ship TODAYfromMILWAUKEE | $356.00 | |
| 1 g | Available to ship TODAYfromMILWAUKEE | $561.00 | |
| 5 g | Available to ship TODAYfromMILWAUKEE | $2,290.00 |
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
Biological source:
Clostridium histolyticum
biological source
Clostridium histolyticum
Quality Level
form
powder
specific activity
≥125 CDU/mg solid, 0.5-5.0 FALGPA units/mg solid
suitability
suitable for release of physiologically active rat epididymal adipocytes
application(s)
diagnostic assay manufacturing
shipped in
wet ice
storage temp.
−20°C
General description
Clostridium histolyticum is a pathogenic clostridium that produces collagenase. This is a mixture of enzymes containing collagenase, non-specific proteases and clostripain.[1]
Application
Collagenase may be used: [1]
- for the preparation of arterial tissue for the study of advanced glycosylation end products (AGE)
- for use along with other proteases for the disaggregation of human tumor, mouse kidney, human brain, lung epithelium and many other tissues.
- in liver and kidney perfusion studies, digestion of pancreas, and isolation of nonparenchymal hepatocytes
- for the preparation of viable hepatocytes from rat liver and for the isolation of fat cells from rat adipose 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.
Collagenase is activated by four gram atom calcium, per mole enzyme. It is inhibited by ethylene glycol-bis(β-aminoethyl ether) - N, N, N′,N′-tetraacetic acid, β-mercaptoethanol, glutathione, thioglycolic acid and 8-hydroxyquinoline.Collagenase enzymes and neutral protease plays an important role in the effective release of cells from tissue. Collagenase recognizes the sequence -R-Pro-8-X-Gly-Pro-R-, where X is most often a neutral amino acid.
Preparation Note
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.
This product also contains clostripain, nonspecific neutral protease, and tryptic activities.
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 | |||
|---|---|---|---|
| biological source Clostridium histolyticum | biological source - | biological source Clostridium histolyticum | biological source Clostridium histolyticum |
| 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 ≥800 CDU/mg solid, 2-5 FALGPA units/mg solid |
| form powder | form powder | form powder | form powder |
| application(s) diagnostic assay manufacturing | application(s) diagnostic assay manufacturing | application(s) diagnostic assay manufacturing | application(s) diagnostic assay manufacturing |
| shipped in wet ice | shipped in - | shipped in wet ice | shipped in - |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
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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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Instructions
C M Posavad et al.
Mucosal immunology, 10(5), 1259-1269 (2017-01-05)
Local mucosal cellular immunity is critical in providing protection from HSV-2. To characterize and quantify HSV-2-reactive mucosal T cells, lymphocytes were isolated from endocervical cytobrush and biopsy specimens from 17 HSV-2-infected women and examined ex vivo for the expression of
O Matsushita et al.
Journal of bacteriology, 181(3), 923-933 (1999-01-28)
Clostridium histolyticum collagenase contains a number of different active components. Previously we have shown that colH encodes a 116-kDa collagenase (ColH) and a 98-kDa gelatinase. We purified a different 116-kDa collagenase (ColG) from the culture supernatant and sequenced its gene
Seglen, P.O.
Methods in Cell Biology, 13 (1976)
Global Trade Item Number
| SKU | GTIN |
|---|---|
| C6885-100MG | 04061835543489 |
| C6885-5G | 04061835543496 |
| C6885-25MG | 04061835556182 |
| C6885-500MG | 04061835556199 |
| C6885-1G | 04061833511664 |
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In case I want to make a stock solution of Collagenase type II of C. histolyticum to store at -20°C, can this be made in PBS or should this PBS already be supplemented with CaCl2? This is not specified in the product information sheet. Thank you.
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