Product introduction

Collagenase I is a black brown powder in appearance. With a molecular weight of 68,000 to 125,000, collagens are able to specifically bind peptide bonds between neutral amino acids (X) and glycine in the -R-Pro-8-X-Gly-pro-R - sequence. The crude product is a mixed enzyme secreted by Clostridium histolytica. They are mainly collagenase, neutral protease and clostridium protein enzyme. Clostridium protease is the least active enzyme that is oxidized, and the separation of tissues must rely on the joint action of collagenase and neutral protease. Activator: Ca2+, the best concentration is 5 mM. Substrate: Different types of collagen are its natural substrates, and some synthetic peptides can also be used as substrates, such as: N-cbz-gly-pro-gly-pro-ala (Km = 0.71 mM), N-CBz-gly-pro-leu-Gly-Pro, N-(3-(2-furyl)acryloyl)- leu-gly-pro-ala(FALGPA), 4-Phenylazo benzyloxycarbonyl-pro-leu-gly-pro-D-arg, n-2,4-Dinitrophenyl- pro-gln-gly-ile-ala-gly-gln-D-arg. In addition, N-Succinyl-gly-pro-leu-glypro7-amido-4-methylcoumarin is a substrate of collagenase-like peptidase. N-(2, 4-dinitrophenyl) -Proleu-gly-Leu-TRp-Ala-D-ARG amino acid compounds are substrates for collagenase in vertebrates.

Scope of application

It is suitable for the isolation of epithelial, lung and adrenal tissue cells.

Usage method

Collagenase I derived from Clostridium tissotica can be formulated as 1-2 mg/ml with TESCA buffer (50 mM TES, 0.36 mM calcium chloride, pH 7.4, 37℃) (Cat:G0150). It can also be prepared with PBS containing calcium and magnesium, hanks containing calcium and magnesium, or serum free medium. After distribution, to avoid repeated freeze-thaw.

Unit definition

1 collagen digestion unit (CDU):at 37℃,pH 7.4 ,and in the presence of calcium ions, polypeptide equivalent to ninhydrin chromogenic 1 micromolar leucine released from collagen every 5 hours is an active unit.

1FALGPA unit: At 25℃, pH 7.5 and in the presence of calcium ions,one FALGPA hydrolysis unit can hydrolyze 1.0μmol of furanyl acrylyl-Leu-gly-pro-Ala per minute.

Optimum pH

6.3-8.8

Inhibitors

EGTA, Reduced Glutathione, β-Mercaptoethanol, Sodium Thioglycolate, 8-Hydroxyquinoline, 2,2' -Bipyridine etc.

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