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Proteinase K (Protease K) is a serine protease with wide cleavage activity, which can cleave the carboxy terminal peptide bonds of aliphatic and aromatic amino acids. Proteinase K has a wide range of applications. It is commonly used in molecular biology, cell biology, immunohistochemistry and other related experiments to digest various proteins, such as preparing chromosomal DNA for pulse electrophoresis, removing nucleases in DNA and RNA preparation, western blotting, Tissue cell permeability and other experiments. Commonly used concentrations of EDTA, Triton X-100, Tween 20, Sarkosyl, and guanidine hydrochloride have little effect on the activity of Proteinase K, while denaturants such as SDS (1%) can increase its activity. The common working concentration of Proteinase K is 50-100 µg /mL, according to whether the buffer used contains SDS, urea, pH, temperature and other factors to determine the specific working concentration.
This product, Recombinant Proteinase K, is a white to off-white freeze-dried powder. It uses the serine protease gene derived from Tritirachium album Limber (Candida albicans), which has been modified by site-directed mutagenesis and recombinantly expressed, purified and lyophilized by Pichia pastoris strains, Does not contain DNase and RNase, the molecular weight is about 30 kDa, and the purity is >95% by SDS-PAGE. The recombinantly expressed mutant Proteinase K has better performance than the wild-type enzyme. It can maintain enzyme activity in a wide pH and temperature range. The effective pH range is pH 7.0-11, and the optimal pH is around pH 10; effective temperature The range is 30-65℃, the best reaction temperature is 55℃, 90% of enzyme activity can be maintained at 50℃-60℃, about 85% of enzyme activity can be maintained at 65℃, and the enzyme activity can be completely inactivated by treatment at 70℃ .
At 37°C and pH 7.5, the amount of enzyme required to release 1 μmol of tyrosine by hydrolysis in 1 minute with casein as a substrate is defined as one unit of enzyme activity. The specific activity of Recombinant Proteinase K produced by our company is >30 U/mg.
Storage and transportation
Transport with wet ice; store at -20°C, valid for 24 months.