Proteinase K

A non-specific serine protease, Proteinase K will inactivate nucleases during native mRNA and DNA preparation. Recommended for high molecular weight nucleic acids from mammalian and microorganism sources. Used in the preparation of genomic DNA from bacteria. Active in the presence of SDS.\n\nSpecific Activity (Protein): \n(same/more than) 30 units/mg dry weight\n\nUnit Definition:\nOne unit liberates 1umole of Folin positive amino acids, measured as Tyrosine, in 10 minutes at 37 degrees C, pH 7.5, using urea-denatured hemoglobin as the substrate.\n\nWorking Concentration:\n3ul of a 20mg/ml solution per 1.5ml of bacterial culture. Dissolve 20mg/ml Proteinase K in 10mM Tris HCl, pH 7.5, 1mM calcium acetate.\n\nOptimum pH: 7.5-12, using denatured hemoglobin as substrate.\n\nSpecificity:\nIn addition to cleavage of peptide bonds, it is able to catalyze peptide amide hydrolysis. Proteinase K is inactivated by diisopropyl fluorophosphate (DFP) or phenyl methane sulphonyl fluoride (PMSF). Chelating agents such as citrate and EDTA have no affect on the enzyme activity.\n\nStorage and Stability:\nLyophilized powder may be stored at 4 degrees C. Stable for 12 months at 4 degrees C. Reconstitute, aliquot and store at -20 degrees C. Reconstituted product is stable for 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.\n\nImportant Note:\nProteinase K solution contains Ca2+ and glycerol and is stable for 6 months. Shipping and short term storage can be at ambient temperature.\n\nAlthough calcium ions do not affect the enzyme activity, they do contribute to its stability when present at a concentration of 1-5 umoles. An interesting characteristic of proteinase K is that it retains its activity in the presence of SDS or urea. (0.5-1% SDS and 1-4 M urea). Raising the temperature of the reaction from 37 degrees C to 50-60 degrees C can increase the activity several fold. A special feature of proteinase K is its ability to digest native proteins, thereby inactivating enzymes such as DNase and RNase without recourse to a denaturation process.

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SPECIFICATIONS

Catalog Number

P9100

Size

100mg

Form

Supplied as a lyophilized powder.

References

US Biological Application Reference: 1. Ellwood, S.R., et al., Genome Biology 11: R109 (2010). General References: 1. Ebeling, W., et al., Eur. J. Biochem, 47: 91-97 (1974). 2. Ausubel, F.M., et al.,

Alternative Names

EC=3.4.21.64; Tritirachium album limber