C0130

Sigma-Aldrich

Collagenase from Clostridium histolyticum

for general use, Type I, 0.25-1.0 FALGPA units/mg solid, ≥125 CDU/mg solid

Synonym(s):
Clostridiopeptidase A
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
NACRES:
NA.54
Pricing and availability is not currently available.

Quality Level

form

essentially salt-free, lyophilized powder

specific activity

≥125 CDU/mg solid
0.25-1.0 FALGPA units/mg solid

mol wt

68,000 to 125,000

optimum pH

6.3-8.8

solubility

TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble 1-2 mg/mL at 37 °C

storage temp.

−20°C

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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. 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. Crude collagenase mixtures contain two major enzyme types namely, collagenase and clostripain.

Application

Collagenase from Clostridium histolyticum has extensive use in biological studies, where it is used to prepare isolated cell suspensions. 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.

Packaging

100, 500 mg in poly bottle
1, 5 g in poly bottle

Biochem/physiol Actions

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.
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

This product is an equivalent to first 40% ammonium sulfate fraction of Mandl, I., et al., J. Clin. Invest., 32, 1323 (1953). 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.

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Personal Protective Equipment

dust mask type N95 (US),Eyeshields,Gloves

Hazard Codes

Xn

Risk Statement

36/37/38-42

Safety Statement

22-24-26-36/37

RIDADR

NONH for all modes of transport

WGK Germany

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

  1. How should I prepare a stock solution of Product C0130, Collagenase?

    Stock solutions of crude Collagenase may be prepared at 10 mg/ml of Collagenase in TESCA buffer (50 mM TES, 0.36 mM calcium chloride, pH 7.4 at 37°C) and are stable if frozen quickly in aliquots and kept frozen at -20°C. Freeze-thaw cycles will damage the enzyme.

  2. How should Product C0130, Collagenase, be prepared for use in tissue digestions?

    For use in tissue culture applications, Collagenase can be solubilized in calcium-free solutions such as Hanks Balanced Salts (H2387) or Earl's' Balanced Salt Solution (E6267).

  3. What is the activity of the lot number of Product C0130, Collagenase, which I have?

    The activity and specific activity for each lot number can be obtained from the certificate of analysis.

  4. What type of Collagenases are present in Product C0130, Collagenase,?

    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. It does not contain any Matrix Metalloprotease.

  5. Is Product C0130, Collagenase, sterile?

    This product does not consist of a sterile preparation of Collagenase. Upon reconstitution, solutions should be sterile filtered through a 0.22 micron filter.

  6. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  7. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  8. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

  9. What is the Department of Transportation shipping information for this product?

    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

  10. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

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