Proteinase K, Molecular Biology Grade (OPGG00001)
|Host||Yeast, Saccharomyces cerevisiae|
|Application||IA, PCR, Molecular Biology, Sample Prep|
|Additional Information||Specific Activity ≥ 34 units/mg of protein. One unit is defined as the enzyme activity that produces 1 µmol of tyrosine per minute from casein at 37°C and pH 7.5.
pI = 8.9 pH Range = 4.5-12.0 (typically used in pH range 7.5-9.0). Maximum activity at 70°C (37-70°C temperature range recommended.)
|::||DNase activity: none detected when 40 µg Proteinase K is incubated with 1 µg λ DNA for 6 hours at 37°C. |
RNase activity: none detected when 40 µg Proteinase K is incubated with 2 µg RNA for 2 hours at 37°C.
|::||Protease and Endotoxin free. |
Note: Proteinase K Molecular Biology Grade (OPGG00001) is NOT sterile. For a sterile product, please refer to item number OPGG00002.
|Reconstitution and Storage||Product is soluble in water at 1 mg/mL. Stock solution can be prepared at 20 - 40 mg/mL in 20 mM Tris-HCl buffer, pH 7.4, and can be sterile filtered using a low protein binding PES or PVDF 0.22 um filter. |
Store the lyophilized format at -20°C up to 2 years.
Enzyme solutions prepared as recommended may be stored at -20°C (with 50% glycerol) for 2 years or at 4°C for one year.
|Protocol Information||Proteinase K is commonly used in molecular biology protocols to remove protein contamination from preparations of highly native undamaged nucleic acid since it rapidly and effectively inactivates nucleases that may degrade the DNA or RNA even in the presence of denaturing reagents. Proteinase K is active in 1% Triton X-100 and fully active in 0.5% (w/v) SDS which denatures protein substrates to increase digestion rates. The enzyme works best at 50-200 ug/mL at pH 7.5-8.0, 37°C and is denatured by subsequent phenol extractions. Incubation times vary from 30 minutes to 18 hours and proteinase K can auto-digest during long incubation. The recombinant enzyme is a mutant engineered for higher specific activity and yield as well as wider pH and temperature activity ranges. The DNAse-free nature of recombinant Proteinase K made it well-suited for isolating PCR and RT-PCR templates.
Activators: 1-5 mM Ca2+ can be added to stimulate proteinase K activity. Optimization using activators can increase proteinase activity significantly. Enzyme activity will be reduced by 25% with the addition of EDTA.
Inhibitors: DIFP or PMSF (5mM). Proteinase K is not inhibited by iodoacetic acid, trypsin-specific inhibitor TLCK, chymotrypsin-specific inhibitor TPCK, and p-chloromercuribenzoate.
|Alias Symbols||PROK;Peptidase K;Endoproteinase K;Endopeptidase K;|
|Protein Name||Proteinase K|
|Protein Accession #||P06873|
|Protein Size (# AA)||384|
|Molecular Weight||29 kDa|
- Reconstitution & Storage Instructions
- Western Blotting/Immunoblotting (WB/IB) Protocol
- Immunohistochemistry (IHC) Protocol
- Immunocytochemistry (ICC) Protocol
- Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol
- Blocking Peptide Competition Protocol (BPCP)
- Immunoprecipitation (IP) Protocol
- Antibody Array (AA) Protocol
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