Collagenase Type 3
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- 适用
- Clostridium histolyticum
- 纯化方法
- Collagenase type 3 is partially purified and contains lowest levels of secondary proteases. The collagenase assay is a modification of the Mandl collagen digestion procedure wherein collagenase is incubated for five hours with native collagen and the extent of collagen breakdown is determined using the Moore and Stein, JBC, 176, 367, (1948) colorimetric ninhydrin method. Amino acids released are expressed as micromoles L-leucine per milligram collagenase in 5 hours at 37°C, pH 7.5. Caseinase activity, a measure of non-specific proteolytic activity, is determined using the above assay and substituting 25 milligrams vitamin free casein for the collagen substrate. Caseinase activity is calculated as for collagenase activity. Clostripain activity is measured after activation in 2.5 mM dithiothreitol (DTT). One unit hydrolyzes one micromole of BAEE per minute at 25°C, pH 7.6, after activation. Tryptic activity is assayed using the same BAEE method as clostripain, but without activation.
- Biological Activity Comment
- Collagenase: ≥100 CDU/mg dry weight Caseinase: ≥50 u/mg dry weight Clostripain: ≤3.0 u/mg dry weight Tryptic: ≤0.3 u/mg dry weight
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CUTANA™ pAG-MNase for ChIC/CUT&RUN Assays
ChIC, CUT&RUN 20 X
Lambda Protein Phosphatase400 U/ µL
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- 应用备注
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Application Note: Collagenase Type 3 is lower in secondary proteolytic contaminant activities but with typical collagenase activity. It is suggested for mammary primary cell isolation. Collagenase is typically used at concentrations from 0.05 % to 0.5 % (w/v) in balanced salt solutions such as Hank's, Earle's and others. For best results the precise mixture of collagenase and proteolytic activities must be tailored to the tissue to be dissociated. Specific conditions for reactivity should be optimized by the end user.
- 说明
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Synonyms: Clostridium histolyticum, Bacterial collagenases, collagenase, caseinase, clostripain, tryptic, ColH, ColG
Background: Crude collagenase preparations contain several isoforms of two different collagenases, a sulfhydryl protease, clostripain, a trypsin-like enzyme, and an aminopeptidase. This combination of collagenolytic and proteolytic activities is effective at breaking down intercellular matrices, the essential part of tissue dissociation. One component of the complex is a hydrolytic enzyme which degrades the helical regions in native collagen preferentially at the Y-Gly bond in the sequence Pro-Y-Gly-Pro, where Y is most frequently a neutral amino acid. This cleavage yields products susceptible to further peptidase digestion. Crude collagenase is inhibited by metal chelating agents such as cysteine, EDTA or o-phenanthroline but not DFP. It is also inhibited by α2-macroglobulin, a large plasma glycoprotein. Ca2+ is required for enzyme activity. Particular enzymatic profiles of each collagenase have been correlated with the tissues from which the cells for study were obtained (or with the uses to which the cells are put) and as a result of the correlations several types of crude collagenases have been established. Crude collagenases are widely used in enzymatic primary cell isolation and tissue dissociation procedures. Most researchers employ either crude collagenase preparations such as Types 1, 2, 3, and 4 or chromatographically purified collagenase, the latter usually combined with secondary enzymes such as elastase, hyaluronidase, etc. Collagenase is ideal for researches focused in Stem Cell and Biomarker Research.
Gene Name: colH, colG
- 限制
- 仅限研究用
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- 状态
- Lyophilized
- 溶解方式
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Reconstitution Volume: 10.0 mL
Reconstitution Buffer: Restore with deionized water (or equivalent)
- 缓冲液
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Buffer: None
Stabilizer: None
- 储存液
- Without preservative
- 储存条件
- 4 °C,-20 °C
- 有效期
- 12 months
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- UniProt
- Q46085
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