Product Number: PK01                           Lyophilized powder     Molecular bio grade

Shipping and Storage

Shipping under room temperature and Store at 4 to -20°C recommended.

Description

Proteinase K is a broad-spectrum serine protease originally isolated from fungus Engyodontium album. Proteinase K is commonly used in molecular biological and biopharmaceutical applications to remove protein contamination from preparations of highly native undamaged nucleic acid because it rapidly and effectively inactive nuclease that might degrade the DNA or RNA even in the presence of denaturing reagents. MEBEP recombinant proteinase K is a mutant to the native protease that result in improved specific activity, higher yield and wider range of pH/temperature with optimal activity. The large scale recombinant preparation has advantages in lot-to-lot consistency, superior purity and cost-efficiency. DNA-free nature of recombinant Proteinase K made it well-suited in isolating DNA and RNA templates. Recombinant proteinase K is widely used for general digestion of proteins and Chemo-enzymatic peptide synthesis in molecular biology, molecular diagnostic and biopharmaceutical applications. Proteinase K is one of important enzymes in various industries like leather, cosmetics, food and feed etc.

Preparation

1. The Molecular Biology Grade lyophilized powder is NOT sterile.
2. Stock solution can be prepared as 40-80 mg/ml in dilution buffer [20mM Tris-HCl (pH 7.4) , 1 mM CaCl₂] or [20mM Tris-HCl (pH 7.4) , 1 mM CaCl₂, 2% Glycerol], sterilized using a 0.22 μm filter and supplied at final concentration of 20-40mg/ml in 50% sterilized Glycerol. Store in aliquots at wide temperature range from 24°C to -80°C.
3. PES or PVDF membranes with low protein binding are recommended in sterile filtration.

Quality Control

1. DNase Activity: none detectable enzyme activity with λ DNA after 6 hours incubation at 37°C.
2. RNase Activity: none detectable of ribonuclease activity after 16 hours incubation at 25°C.
3. Protein Purity: > 95% (Native-PAGE and SDS-PAGE assay).

Description

Storage Conditions

Store at 4 to -20ºC recommended.
Mutated Proteinase K powder is stable at room temperature in 3 years.
Expiration: Three years if kept below 4ºC.

Dilution Buffer 1 (short time storage): 20 mM Tris-Cl (pH 7.4), 1 mM CaCl2, 15% glycerol, or d3H2O.
Dilution Buffer 2 (long term storage): 20 mM Tris-Cl (pH 7.4), 1 mM CaCl2, 50% glycerol, 0.02-0.1% Sodium azide.

QC Assays

• Activity Assay and Unit Definitions: One unit is defined as the amount of enzyme that will liberate 1 μmol of tyrosine per minute at 37º C, pH7.5.
• DNase Activity: none detectable enzyme activity with λ DNA after 6 hours incubation at 37ºC.
• RNase Activity: none detectable of ribonuclease activity after 16 hours incubation at 25 º C
• Protein Purity: over 99% (Native-PAGE and SDS-PAGE assay)

Applications

The application of this Mutated Proteinase K is similar with wild type Proteinase K. But the mutated one has higher specific activity and more stable at RT. It is a broad substrate non-specific serine Proteinase. It is active over the pH range 4-11. It was used on any situation to digest native and denatured proteins. For instance, it is used for isolating mRNA or genomic DNA from different tissues and modifying glycoprotein for structure studies. Mutated Proteinase K is active with SDS, urea and EDTA and active between the temperature15-75ºC

Other Names

• Proteinase K
• endopeptidase K
• Tritirachium alkaline proteinase
• Tritirachium album proteinase K
• Tritirachium album serine proteinase

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