Recombinant Proteinase K (Powder) 1g

ProductDescription/Introduction

Proteinase K is a broad spectrum protease. Its predominant cleavage site is the peptide bond adjacent to the carboxyl group of aliphatic and aromatic amino acids with blocked α-amino groups. Proteinase K is widely used, 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 permeabilizing 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 the SDS (1%) can increase its activity. The usual working concentration of Proteinase K is 50-100 µg/mL, and the specific working concentration is determined according to whether the buffer containing the SDS, urea and the pH, temperature of the buffer.

    The gene of this recombinant Proteinase K is from Tritirachium album Limber, was mutated by site-directed. Mutant is expressed in Pichia pastoris, purified, does not contain DNase, RNase. The molecular weight is about 30kDa. The purity is ≥95% by SDS-PAGE. This recombinant mutant of Proteinase K has better performance than the wild-type, and it can maintain enzyme activity in a wide pH and temperature range. The effective pH range is pH 7.0-11, with optimal activity at around pH 10. The effective temperature range is 30-65℃. The optimal temperature is 55℃, and 80% of enzyme activity can be maintained at 50℃-60℃, and about 85% of enzyme activity can still be maintained at 65℃. The enzyme will be completely inactivated at 70℃. One unit of enzymatic activity was defined as the amount of proteinase K needed to catalyze the release of 1 μmol of tyrosine per minute at 37℃ and pH 7.5. The specific activity of the Proteinase K produced by our company is ≥30 U/mg.

Storage andShipping Conditions

Ship with wet ice; Store at -20℃ valid for 24 months.

Assay Protocol / Procedures

Resuspend at a stock concentration that is convenient for your experimental needs. (Dissolve in 20 mM Tris HCl, pH 7.5, 50 mM Sodium chloride , 5 mM calcium chloride, and 50% glycerol, then store at –20°C)

Note

1. It is recommended to use in sub- package to avoid freeze -thawing cycles.

2. Due to the high activity and stability of proteinase K, cross-contamination should be avoided.

3. For your safty and health,please wear safety glasses, gloves, or protective clothing.

For Research Use Only!