All Photos(3)

C6885

Sigma-Aldrich

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
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
NACRES:
NA.54

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

Featured Industry

Diagnostic Assay Manufacturing

shipped in

wet ice

storage temp.

−20°C

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General description

Clostridium histolyticum is a pathogenic clostridium that produces collagenase. This is a mixture of enzymes containing collagenase, non-specific proteases and clostripain.

Application

Collagenase may be used:
  • 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

Packaging

100, 500 mg in poly bottle

Biochem/physiol Actions

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.
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.

Caution

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.

Unit Definition

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

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.

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Precautionary Statements

Hazard Classifications

Resp. Sens. 1

Storage Class Code

11 - Combustible Solids

WGK Germany

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US),Eyeshields,Gloves

Certificate of Analysis

Certificate of Origin

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...
Seglen, P.O.
Methods in Cell Biology, 13 (1976)
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...
Methods in Enzymology, 192, 829-829 (1990)
D. Schomberg and M. Salzmann
Enzymes, 1 (1991)

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