Selection Guide for Collagenase for Animal Tissue Digestion
Product Introduction
Collagenase is a type of endopeptidase that can specifically recognize the Pro-X-Gly-Pro sequence (this sequence appears 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. It can also hydrolyze collagen in connective tissue.
Many proteases can hydrolyze single-chain and denatured collagen polypeptides, but collagenase is the only protease that can degrade natural collagen fibers with a triple-stranded superhelical structure. It is widely present in connective tissue. Among them, the effective collagenase is the "crude " collagenase secreted by the anaerobic bacterium Clostridium histolyticum .
Yisheng collagenase is derived from Clostridium histolyticum and is a "crude" enzyme extract. It not only contains collagenase that can degrade natural collagen and reticular fibers, but also contains other proteases such as polysaccharidase, lipase, etc., which can effectively hydrolyze other proteins, polysaccharides and lipids in the extracellular matrix of connective tissue and epithelial tissue, and is therefore more suitable for tissue digestion.
Currently, bacterial collagenase on the market is mainly divided into five types based on the differences in collagenase activity: collagenase type I, type II, type III, type IV and type V.
Many proteases can hydrolyze single-chain and denatured collagen polypeptides, but collagenase is the only protease that can degrade natural collagen fibers with a triple-stranded superhelical structure. It is widely present in connective tissue. Among them, the effective collagenase is the "crude " collagenase secreted by the anaerobic bacterium Clostridium histolyticum .
Yisheng collagenase is derived from Clostridium histolyticum and is a "crude" enzyme extract. It not only contains collagenase that can degrade natural collagen and reticular fibers, but also contains other proteases such as polysaccharidase, lipase, etc., which can effectively hydrolyze other proteins, polysaccharides and lipids in the extracellular matrix of connective tissue and epithelial tissue, and is therefore more suitable for tissue digestion.
Currently, bacterial collagenase on the market is mainly divided into five types based on the differences in collagenase activity: collagenase type I, type II, type III, type IV and type V.
Selection of Collagenase
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*CDU = collagen digestion units
Product Features
1. High digestion efficiency: The crude enzyme extract of Clostridium histolyticum has high efficiency in tissue digestion.2. Dependence on activators: Collagenase requires the help of divalent cations such as Ca 2+ and Zn 2+ to activate the activity of the enzyme.3. Affected by inhibitors: The inhibitors of collagenase include EDTA, EGTA, etc.; the second is heavy metal ions (Hg2 + , Pb2 + , Cd2 + ); and some chemical reagents such as Cysteine, histidine, DTT, 2-mercaptoethanol, o-phenanthroline, etc. also have inhibitory effects on collagenase.4. Strong resistance to interference: Collagen will not be affected by the serum used to culture cells.
FAQ
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℃
3. Suggestions for use:① For the study of collagen structure and biosynthesis, higher purity collagenase preparations are often used, which do not contain other proteolytic activities.②For tissue dissociation, crude collagenase preparations such as elastase, trypsin or papain are often used.
4. Poor digestion during the experiment① If the enzyme activity is weak or calcium ion is insufficient, collagenase should be stored under low temperature and dry conditions, and the collagenase solution should be divided and frozen.②If it is caused by insufficient calcium ions, the calcium ion concentration in the collagenase solution should be set to 5mM.
5. Cell death occurs during use① If there is an overdose of protease, the protease contact time should be reduced or albumin or inactivated serum should be added.②If the pH changes , use a buffer solution (such as HBSS ); check and readjust the pH frequently.③ If there is too little oxygen, sterile air should be passed through the digestive fluid
6. Cell damage during the experimentIf the cause is excessive protease, the amount of protease should be reduced or albumin or inactivated serum should be added.
4. Poor digestion during the experiment① If the enzyme activity is weak or calcium ion is insufficient, collagenase should be stored under low temperature and dry conditions, and the collagenase solution should be divided and frozen.②If it is caused by insufficient calcium ions, the calcium ion concentration in the collagenase solution should be set to 5mM.
5. Cell death occurs during use① If there is an overdose of protease, the protease contact time should be reduced or albumin or inactivated serum should be added.②If the pH changes , use a buffer solution (such as HBSS ); check and readjust the pH frequently.③ If there is too little oxygen, sterile air should be passed through the digestive fluid
6. Cell damage during the experimentIf the cause is excessive protease, the amount of protease should be reduced or albumin or inactivated serum should be added.