Product introduction

Collagenase II is a crude product and appears as a black brown powder. Molecular weight 68,000 to 125,000, optimal PH 6.3-8.8. Collagenase can specifically bind peptide bonds between the neutral amino acid (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 were mainly collagenase, neutral protease and clostridium protease. 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+, each mole of enzyme requires 4g calcium ions to activate. 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 adipose, liver, bone, thyroid, heart and salivary gland tissue cells, especially epithelial tissue.

Usage method

Collagenase derived from Clostridium tissotica collagenase II 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℃, pH7.4 and in the presence of calcium ions, the amount of polypeptide equivalent to ninhydrin chromogenic 1 micromolar leucine released from collagen every 5 hours was taken as one active unit. 1FALGPA unit: At 25℃, pH7.5 and in the presence of calcium ions, one FALGPA hydrolysis unit can hydrolyze 1.0μmol of furanyl acrylyl-LeU-gly-pro-ALA per minute.

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