产品名称

Proteinase K from Tritirachium album

别 称

Endopeptidase K

含 量

≥800units/mL

CAS  NO.

39450-01-6

分子量

mol wt 28.93 kDa

MDL  number

MFCD00132129

适用范围

生物试剂,  for molecular biology

l  理化信息

外 观

清撤至非常朦胧的无色至浅黄色至浅棕色缓冲的甘油水溶液

物理形态

Solution in 40% glycerol (v/v) containing 10mM Tris-HCl, pH7.5, with 1mM calcium acetate.

l  生物学信息

生物活性/药理作用

Proteinase K is a stable and highly reactive serine protease. Evidence from crystal and molecular structure studies indicates the enzyme belongs to the subtilisin family with an active-site catalytic triad (Asp39-His69-Ser224). It is stable in a broad range of environments: pH, buffer salts, detergents (SDS), and temperature. In the presence of 0.1-0.5% SDS, proteinase K retains activity and will digest a variety of proteins and nucleases in DNA preparations without compromising the integrity of the isolated DNA.

Proteinase K has a broad specificity and degrades many proteins even in the native state. It mainly cleaves the peptide bond adjacent to the carboxyl group of aliphatic and aromatic amino acids with blocked α-amino groups. The molecular weight of proteinase K from amino acid sequence is found to be 28,930 Da and from SDS-PAGE, it is found to be 28,500 Da. The optimum pH is between 7.5-9.0 and its isoelectric point is 8.9. Ca₂₊ (1-5mM) is required for its activation. Proteinase K is inhibited by DIFP (diisopropylfluorophosphate) or PMSF(phenylmethylsulfonyl fluoride).

应 用

The product has been used to study its pre-treatment effects on the silk fibroin. The aspects analysed in this study included the crystallographic properties of hydroxyapatite (HAp), and the microstructure and microhardness of the composites. The enzyme has also been used to facilitate the access of probes to rRNA using FISH techniques to detect pathogenic Staphylococcus aureus.

Useful for the proteolytic inactivation of nucleases during the isolation of DNA and RNA.

Removes endotoxins that bind to cationic  proteins such as lysozyme and ribonuclease A.

Reported useful for the isolation of hepatic, yeast, and mung bean mitochondria

Determination of enzyme localization on membranes

Treatment of paraffin embedded tissue sections to expose antigen binding sites for antibody labeling.

Digestion of proteins from brain tissue samples for prions in Transmissible Spongiform Encephalopathies (TSE) research.

样品准备说明

Solution in 40% glycerol (v/v) containing 10mM Tris-HCl, pH7.5, with 1mM calcium acetate.

有效单位定义

One unit will hydrolyze urea-denatured hemoglobin to produce color equivalent to 1.0μmole of tyrosine per min at pH7.5 at 37°C (color by Folin-Ciocalteu reagent).

l  存储

存储温度

2-8°C

l  注意事项及免责声明

本产品仅用于实验研究，不得作为药物使用，不得用于家用或其它用途。

l  参考文献

1.    http://www.drugbank.ca

2.    https://ncit.nci.nih.gov

3.    https://www.ncbi.nlm.nih.gov