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Cat. No.: CCH2-25 (for 25mg)

Cat. No.: CCH2-100 (for 100mg)

Description

Collagenase is a protease that cleaves a protein called collagen, which is a triple-helical protein. There are three types of tissue collagenases, all of which belong to the matrix metalloproteinase (MMP) family. Collagenase obtained from Clostridium histolyticum has very high activity because it can digest collagen from both ends at temperatures as low as 4-10°C. The crude collagenase mixture contains two major enzyme types, namely collagenase and clostripain.

Specifications    

Reagent grade, Type II, powder, ≥125 CDU/mg solid

Application

Collagenase derived from Clostridium histolyticum has a wide range of applications in biological research and can be used for the preparation of isolated cell suspensions. This product is suitable for the disaggregation of human tumors, mouse kidneys, adult and fetal human brains, lungs, and many other epithelial tissues. It has also been proven effective in liver and kidney perfusion studies, pancreatic digestion, isolation of non-parenchymal rat liver cells, and liver cell preparation. Collagenase has been used in the preparation of arterial tissues required for advanced glycation end-product research. The enzyme has been tested to release liver cells at a concentration of approximately 1 mg/mL. The concentration range for digestion is typically 0.1 to 5 mg/mL.

Biochem/physiol Actions

The collagenase product is a mixture of enzymes secreted by Clostridium histolyticum, and different products can be distinguished by the ratio of 10-18 compounds found in the secreted enzymes. The main components are two types of collagenase, clostripain, and a neutral protease. These enzymes work in synergy to degrade collagen and other intracellular substances. The action of collagenase and the neutral protease is necessary for the effective release of cells from tissues. Different types of collagen are natural substrates for collagenase. Each mole of collagenase is activated by 4g of atomic calcium. It is inhibited by ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid, β-mercaptoethanol, glutathione, mercaptoacetic acid, and 8-hydroxyquinoline.

Caution

According to the provided information, this product can be stably stored for one year at -20°C. Activity of FALGPA or protease is not lost within 30 days at 37°C, 50°C, and -20°C. If aliquoted (10mg/mL) and rapidly frozen at -20°C, the solution of crude collagenase is stable. Further repeated freeze-thaw cycles will damage the solution. This product can maintain 100% activity when kept on ice for over 7 hours.

Preparation Note

This product is equivalent to the original 40% ammonium sulfate fraction [Mandl, I., et al., J. Clin. Invest., 32, 1323 (1953)]. The solution is 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).

Unit Definition

• One Collagen Digesting Unit (CDU) releases an amount of peptide chains from bovine Achilles tendon collagen equivalent to 1.0 μmole leucine, in 5 hours, at pH 7.4, 37°C, in the presence of phenylmethylsulfonyl fluoride (PMSF).
• One FALGPA hydrolysis unit hydrolyzes 1.0 μmole of N-α-benzoyl-L-arginine ethyl ester (BAEE) per minute at 25°C.
• One Neutral Protease Unit hydrolyzes casein to produce color equivalent to 1.0 μmole of tyrosine every 5 hours at pH 7.5, 37°C.
• One Clostripain Unit hydrolyzes 1.0 μmole of BAEE per minute at pH 7.6, 25°C in the presence of dithiothreitol (DTT).

Only for research and not intended for treatment of humans or animals

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