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C9722

Sigma-Aldrich

Collagenase from Clostridium histolyticum

lyophilized powder (from 0.2μm filtered solution), 0.5-5.0 FALGPA units/mg solid, suitable for cell culture

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Synonym(s):
Clostridiopeptidase A
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
NACRES:
NA.75

biological source

Clostridium histolyticum

Quality Level

sterility

0.2 μm filtered

form

lyophilized powder (from 0.2μm filtered solution)

specific activity

0.5-5.0 FALGPA units/mg solid

concentration

1 mg/mL

technique(s)

cell culture | mammalian: suitable
single cell analysis: suitable

pH

7.4

shipped in

ambient

storage temp.

−20°C

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This Item
C9697C2674C2799
sterility

0.2 μm filtered

sterility

0.2 μm filtered

sterility

-

sterility

-

form

lyophilized powder (from 0.2μm filtered solution)

form

lyophilized powder (from 0.2 μm filtered solution)

form

lyophilized powder

form

powder

specific activity

0.5-5.0 FALGPA units/mg solid

specific activity

≥800 units/mg solid

specific activity

≥125 CDU/mg solid (CDU = collagen digestion units), 0.5-5.0 FALGPA units/mg solid

specific activity

≥4 FALGPA units/mg solid, ≥700 CDU/mg solid (CDU = collagen digestion units)

concentration

1 mg/mL

concentration

1 mg/mL

concentration

1 mg/mL

concentration

-

technique(s)

cell culture | mammalian: suitable, single cell analysis: suitable

technique(s)

cell culture | mammalian: suitable

technique(s)

cell culture | mammalian: suitable, single cell analysis: suitable

technique(s)

cell culture | mammalian: suitable

Application

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 1 mg/mL. Concentrations for digestion range from 0.1 to 5 mg/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.

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

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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Protocols

Enzymatic Assay of Collagenase (EC 3.4.24.3) using FALGPA (N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala)

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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