蛋白酶 K ( Proteinase K ),从林伯氏白色念球菌( tritirachium album limber )中提取,是一种切割活性较广的丝氨酸蛋白酶。它切割脂族氨基酸和芳香族氨基酸的羧基端肽键,常用于生物样品中蛋白质的一般降解。该产品经过纯化,去除了 RNase 和 Dnase 活性。由于蛋白酶 K 在尿素和 SDS 中稳定,还具有降解天然蛋白质的能力,因而它应用很广泛,包括制备脉冲电泳的染色体 DNA ,蛋白质印迹以及去除 DNA 和 RNA 制备中的核酸酶。蛋白酶 K 的一般工作浓度是 50 — 100μg/ml 。
配方:将蛋白酶 K 溶于 10mM 的 Tris-HCl ( pH 为 7.5 )、 1mM 氯化钙和 50 %甘油中, 以制成 20mg/ml 的蛋白酶 K 溶液。
Proteinase K, produced by the fungusTritirachium albumLimber, is a serine protease that exhibits broad cleavage activity. It cleaves peptide bonds adjacent to the carboxylic group of aliphatic and aromatic amino acids and is useful for general digestion of protein in biological samples. It has been purified to remove RNase and DNase activities. The stability of Proteinase K in urea and SDS and its ability to digest native proteins make it useful for a variety of applications including preparation of chromosomal DNA for pulsed-field gel electrophoresis, protein fingerprinting and removal of nucleases from preparations of DNA and RNA. A typical working concentration for Proteinase K is 50–100μg/ml.
Formulation:Proteinase K (PK) Solution is supplied at a concentration of 20mg/ml in 10mM Tris-HCl (pH 7.5), 1mM calcium chloride and 50% glycerol.
Features - Benefits
Stable:Active over a pH range of 4.3–12.0 in 0.5% SDS or 1% Triton® X-100 and retains >80% of its activity at temperatures up to 60°C.
Easy to Use:Provided in solution stable at room temperature and does not require resuspension or thawing prior to use.
References
Ebeling, W.et al.(1974)Eur. J. Biochem.47, 91–7.
Schwartz, D.C. and Cantor, C.R. (1984)Cell37, 67–75.