产品名称

Proteinase K from Tritirachium album

别 称

Endopeptidase K

含 量

≥500units/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

l  理化信息

外 观

缓冲甘油水溶液

物理形态

Solution in 40% (v/v) glycerol 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 alpha-amino groups.The optimum pH is between 7.5-9.0and the isoelectric point is 8.9. Ca₂₊(1-5mM) is required for activation. Proteinase K is inhibited by DIFP or PMSF.

应 用

Product has been used to break down cardiac muscle during histopathology studies. It has also been used during the digestion of HEK-293 cells.

The enzyme has been used in the digestion of sealed cytosolic side out ER vesicles. It has been used to deproteinize dissected brain and/or whole pupae sections of honey bee prior to in situ hybridisation. This was done during the study of neuropeptide Y-like signaling, and nutritionally-mediated gene and behaviour in the honey bee.

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% (v/v) glycerol containing 10mM Tris-HCl, pH7.5, with 1 mM calcium acetate. Proteinase K in solution is stable over a pH range of 4.0-12.5 (optimum pH8.0), and is  also stable over the temperature range of 25°C to 65°C during use. At pH 8.0, solutions will be stable for at least 12 months at 4°C. At pH4-11.5, solutions containing Ca₂₊ (1-6 mM) are expected to be stable for several weeks. An 80% ammonium sulfate suspension stored at 4°C is stable for at least 12 months.

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