Proteinase Okay is a broad-spectrum serine protease (28.9kDa monomer) that cleaves peptide bonds on the carboxylic sides of aliphatic, fragrant, and hydrophobic amino acids.
Description
Proteinase Okay is a broad-spectrum serine protease (28.9kDa monomer) that cleaves peptide bonds on the carboxylic sides of aliphatic, fragrant, and hydrophobic amino acids.
Functions
Isolation of genomic DNA from cultured cells and tissues, elimination of DNAses and RNAses throughout DNA and/or RNA purification, dedication of enzyme areas
Extra data
Options
Amount
100 mg lyophilized powder purified from purified from Pichia pastoris harbouring the gene encoding endolytic protease from Tritirachium
album.
Specs
Freed from DNAses and RNAses. Particular Exercise ~30 Kunitz items per mg.
Storage
For brief time period, retailer lyophilized powder at +4ºC. Earlier than use, reconstitute Proteinase Okay at a desired focus utilizing both ultrapure water or 50mM Tris-HCl pH 7,5 containing 5mM CaCl2 and 50% glycerol. Reconstituted Proteinase Okay needs to be saved at -20ºC.
Enzyme exercise
Proteinase Okay is activated by calcium (1-5mM). Though Ca2+ doesn’t immediately influences catalytic exercise it does contributes to the protein stability (safety towards autolysis, growing thermal stability). Elimination of Ca2+ (e.g. by including EDTA) reduces proteolytic exercise by 80%, nevertheless, the residual exercise is adequate to digest proteins, which normally contaminate nucleic acid preparations. Due to this fact, the digest with Proteinase Okay for the purification of nucleic acids is carried out within the presence of EDTA (inhibition of magnesium-dependent enzymes similar to DNAses). Proteinase Okay can also be secure over a large pH vary (4-12), with a pH optimum of
pH 8.0. Elevation of the response temperature from 37°C to 50 – 60°C could enhance the exercise a number of instances, as may the addition of 0.5 – 1% SDS, 3M of Guanidinium chloride, 1Mof Guanidinium thiocyanate, and 4M urea. The beneficial working focus of Proteinase Okay is 0,05-1 mg/ml.
Inhibition
Proteinase Okay may be inhibited by phenylmethylsulfonyl fluoride (PMSF), trichloroacetic acid (TCA), 4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF), and diisopropyl phosphorofluoridate (DFP or DIFP). Proteinase Okay is NOT inhibited by metallic chelators, thiol-reactive reagents, or by particular trypsin or chymotrypsin inhibitors (together with SDS, Tween-20, Triton X-100, urea, EDTA, citrate, iodoacetic acid, Sarkosyl, Guanidinium chloride, Guanidinium thiocyanate, TLCK, and TPCK)
Inactivation
Proteinase Okay may be heat-inactivated at temperatures above 65°C
Paperwork
Obtain Product Information Datasheet
Obtain MSDS GE010 Proteinase Okay v7E502
Order Info – “what is protein k”
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