Application Notes | Nucleic Acids Isolation, Immunohistochemistry, Endotoxin Removal, Nucleases Removal, Protein Contamination Removal |
Comment |
Inhibitor: The enzyme is inactivated by DIFP or PMSF (PMSF used at final concentration 5 mM. ). However, it is not inhibited by EDTA , iodoacetic acid, trypsin-specific inhibitor TLCK, chymotrypsin-specific inhibitor TPCK, and p-chloromercuribenzoate. |
Protocol |
Proteinase K is commonly used in molecular biology to remove protein contamination from preparations of highly native undamaged nucleic acid because it rapidly and effectively inactives nuclease that might degrade the DNA or RNA even in the presence of denaturing reagents. Proteinase K is active in 1% Tritonä 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 mg/mL at pH 7.5-8.0, 37 °C and is usually denatured by subsequent phenol extractions. Incubation times vary from 30 minutes to 18 hours and proteinase K can auto-digest during long incubation. |
Restrictions | For Research Use only |
Format | Liquid |
Concentration | 20 mg/mL |
Buffer | 20 mM Tris-HCl, 1 mM CaCl2, 50 % Glycerol, pH 7.4 |
Storage | 4 °C |
Storage Comment | 4°C |
Supplier Images |
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