Recombinant proteinase K is extremely efficient on native proteins
Calcium is important for the thermostability of proteinase K and the pH value is critical for high solubility
The use of cryoprotectant glycerol to a final concentration of 50% in the stock solution is to prevent freeze / thaw damage. Proteinase K retained 99% of the activity after 12 freeze / thaw cycles in the stock solution. Avoid repeated freezing and thawing to avoid protein precipitation if glycerol is not added.
Proteinase K solution remains active in a wide pH range (4.0-12.5, often used in the 7.5-9.0 range) and in a wide temperature range of 25-75 ° C. The solution remains sterile and stable for at least 12 months at 4 ° C.
Recombinant proteinase K is a nonspecific serine protease that is useful for general protein digestion.
Proteinase K is a highly reactive serine protease that shows the ability to digest native proteins, thus inactivating enzymes such as DNase and RNase without resorting to a denaturation process. It is the most potent proteinase among all the proteinases characterized so far. It cleaves at the peptide bond adjacent to the carboxylic acid group of aliphatic, aromatic, or hydrophobic amino acids.
Recombinant proteinase K is used in the isolation or preparation of high molecular weight nucleic acids. It is highly pure and has a higher specific activity and is more stable at room temperature compared to native proteinase K.
Recombinant proteinase K is expressed from the yeast Pichia pastoris with a cloned gene encoding the endolytic protease of Engyodontium album (Tritirachium album).
Applications
• Recombinant proteinase K is extremely efficient on native proteins and therefore can be used to rapidly inactivate endogenous nucleases such as RNases and DNases. This property makes Proteinase K particularly suitable for the isolation of native RNA and DNA from tissues or cell lines.
• It is used for the analysis of membrane structures by modifying proteins and glycoproteins on the cell surface.
• Particularly suitable for isolating nucleic acids for amplification reactions.
• Promotes cell lysis by activating a bacterial autolytic factor.
• Used to remove cell debris during colony lift preparation and to treat tissue sections to ensure efficient probe infiltration during in situ hybridization.
Reconstitution Recommendations
For the preparation of the stock solution, we suggest reconstituting the lyophilized powder in a buffer (20 mM Tris, 1 mM CaCl2, 2% glycerol, pH 7.4).