Product Introduction

Collagenase is a class of endonucleases that can specifically recognize the Pro-X-Gly-Pro sequence (which occurs frequently in collagen and is rarely found in other proteins), and cleave the peptide bond between the neutral amino acid (X) and glycine (Gly) in this sequence, and can hydrolyze collagen in connective tissue.

Many proteases are capable of hydrolyzing single-stranded and denatured collagen peptides, but collagenase is the only protease that can degrade natural collagen fibers with a three-strand supercoiled structure, and is widely found in connective tissues, the most potent of which is the "crude" collagenase secreted by the anaerobic bacterium Clostridium histolyticum.

Yeasen collagenase is derived from Clostridium histolyticum, which is an enzyme "coarse" extract, which not only contains collagenase that can degrade natural collagen and reticular fibers, but also contains some other proteases such as polysaccharidases, lipases, etc., which can effectively hydrolyze other proteins, polysaccharides and lipids in the extracellular matrix of connective tissue and epithelial tissue, so it is more suitable for tissue digestion.

At present, the bacterial collagenase on the market is mainly divided into five types according to the difference in the activity of collagenase: collagenase type I, type II, type III, type IV and type V.

Choice of collagenase

*CDU = collagen digestion units

Product Features

1. High digestion efficiency: the crude enzyme extract of Clostridium histolyticum has high digestion efficiency for tissues.
2. Dependent on activators: collagenase needs to rely on the activity of activating enzymes such as divalent cations such as Ca²⁺and Zn²⁺.
3. Affected by inhibitors: inhibitors of collagenase include EDTA, EGTA, etc.; the second is heavy metal ions (Hg²⁺, Pb²⁺, Cd²⁺); There are also some chemical agents such as Cysteine, histidine, DTT, 2-mercaptoethanol, o-phenanthroline, etc., which also have inhibitory effects on collagenase.
4. Strong interference resistance: collagen will not be affected by the serum used to culture cells.

Precautions for use

1. Usage: Dissolve in Hank's, Earle's or other balanced salt solutions, the recommended working concentration is 0.05% ~0.5% (w/v)
2. Storage temperature: 2-8°C
3. Suggestions for use:

(1) For the study of collagen structure and biosynthesis, higher purity collagenase preparations are often used, and no other proteolytic activities are used.

(2) For tissue dissociation, crude collagenase preparations, such as elastase, trypsin or papain, are often used.

FAQ

1. Poor digestion during the experiment

(1) If it is caused by weak enzyme activity or insufficient calcium ions, collagenase should be stored under low temperature and dry conditions, and collagenase solution should be frozen after aliquoting

(2) If it is caused by insufficient calcium ions, the concentration of calcium ions in the collagenase solution should be 5mM

2. Cell death occurs during use

(1) If there is an excess of protease, the protease contact time should be reduced or albumin or inactivated serum should be added

(2) If the pH changes, a buffer (e.g., HBSS) should be used; Check and readjust your pH frequently

(3) If there is too little oxygen, the digestive solution should be ventilated with sterile air

3. Cells were damaged during the experiment

 If it is caused by too much protease, the amount of protease should be reduced or albumin or inactivated serum should be added

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