Worthington collagenase raw material

AI Meijie Worthington Collagen enzyme raw material certification ：

Crude collagenase preparation contains several isoforms of two different collagenases , A sulfhydryl protease 、 Clostridium protease 、 A trypsin like enzyme and an aminopeptidase . This combination of collagen hydrolysis and proteolytic activity can effectively decompose the intercellular matrix , This is an important part of organizational dissociation . One component of the complex is a hydrolase , It takes precedence over the sequence Pro-Y-Gly-Pro Medium Y-Gly Degradation of helical regions in natural collagen at the bond , among Y The most common are neutral amino acids . This cleavage produces products that are sensitive to further peptidase digestion . Crude collagenase is replaced by cysteine 、EDTA Or metal chelating agents such as o-phenanthroline inhibit , but DFP Do not suppress . It is also by a large plasma glycoprotein α2- Macroglobulin inhibits . calcium 2+ It is necessary for enzyme activity . The specific zymogram of each collagenase has been compared with the tissue from which the study cell was obtained （ Or with the purpose of placing cells ） relevant , And as a result of correlation ,Worthington Several types of crude collagenases have been established ： type 1、2、3 and 4.

• 1 Type crude collagenase has collagenase 、 Casein enzyme 、 The original balance of Clostridium protease and trypsin activity .

• 2 Type A contains a relatively high level of protease activity , Especially Clostridium protease .

• 3 Type A contains the lowest level of secondary protease .

• 4 Type A is designed to have particularly low trypsin activity , To limit the damage of proteins and receptors .

• 5 type It contains high collagenase and casein enzyme values .

• 6 type Contains high collagenase activity , The ratio of casein enzyme to collagenase is about 2:1. Designed relative to I type ( col  G) Rich in II type ( col  H) Collagenase .

• 7 The activity ratio of type I collagenase and casein enzyme 1 The type and 2 Type I collagenase is four times higher .

AI Meijie Worthington Non animal type AFA、AFB and AFC、STZ1 and STZ2 Worthington  A culture derived from a medium completely free of animal ingredients , Designed for use in bioprocessing applications where the introduction of potential pathogens of animal origin must be prevented . The level of secondary protease is related to 1 The type and 2 Type a collagenase is similar .

•  CLSAFA It's the original AFA Grade , Intended to contain a substance similar to 1 The type and 2 Collagenase and secondary protease of type a collagenase .

•  CLSAFB Content ratio CLSAFA Higher collagenase and caseinase activity .

•  CLSAFC It has a particularly low trypsin activity , Be similar to 4 Type collagenase .

•  STZ1 and STZ2,0.22μ The filter STEMxyme AOF Collagenase / Neutral protease (Dispase) Mixture , It is used to separate primary cells and stem cells .

The collagenase assay is Mandl Improvement , Collagenase is incubated with natural collagen 5 Hours , And use Moore and Stein, JBC, 176 , 367, (1948) The degree of collagen decomposition was determined by colorimetric ninhydrin method . The released amino acids are expressed in micro moles of leucine per mg of collagenase .

AI Meijie Worthington Collagenase uses ：

Crude collagenase is widely used in enzymatic primary cell separation and tissue dissociation procedures . Most researchers either use crude collagenase preparations , for example 1、2、3 and 4 type , Either use chromatographic purified collagenase （ Code ：CLSPA）; The latter is usually associated with secondary enzymes such as elastase 、 Hyaluronidase, etc . For best results , The precise mixing of proteolytic activity must be adjusted for the tissue to be dissociated . There is a good correlation between the types and effectiveness of different organizations , But it's not perfect , And may depend in part on the parameters and objectives used and the differences between batches .