Proteinase K, recombinant, PCR Grade
from Titrirachium album, expressed in Pichia pastoris,
lyophilizate
Application Proteinase K, recombinant, PCR Grade, digests native proteins very efficiently.
This enzyme can be used to rapidly inactivate endogenous RNases and
DNases during nucleic acid isolation. Proteinase K is particularly suited for the
isolation of native RNA and DNA from tissues and cell lines.
The enzyme promotes cell lysis by activating a bacterial autolytic factor.
Proteinase K is also used for:
■ Analysis of membrane structures by modifying proteins and glycoproteins
on cell surfaces
■ Removal of cellular debris during the preparation of colony lifts
■ Treatment of tissue sections to ensure efficient probe infiltration during in
situ hybridization
Properties Nomenclature: Proteinase K
Molecular weight: 28.8 kD
pH optimum: 7.5-10.5
Inhibitors: Proteinase K is inhibited by diisopropyl fluorophosphate and phenylmethylsulfonyl
fluoride (PMSF) and is also totally inactivated by mercuric
ions. Pefabloc® SC and Pefabloc® PLUS are specific, irreversible and nontoxic
inhibitors.
Specificity: Proteinase K is one of the most active endopeptidases known and
does not exhibit any pronounced cleavage specificity. Activity can be stimulated
by addition of denaturing agents (SDS and urea).
Specification Appearance: White lyophilizate
Solubility: Clear, colorless solution in water (c=20 mg/ml)
Volume activity (+37°C, hemoglobin): ≥24 U/mg lyophilizate
Specific activity (+37°C, hemoglobin): ≥30 U/mg protein
Unit definition (hemoglobin): One unit is the enzyme activity which releases
folin positive amino acids and peptides equivalent to 1 μmol of tyrosine in 1
minute under the test conditions.
Volume activity (+25°C, Chromozym): ≥2 U/mg lyophilizate
Specific activity (+25°C, Chromozym): ≥2.5 U/mg protein
Unit definition (Chromozym): One unit is the enzyme activity which cleaves at
+25°C in 1 minute 18 mmol Chromozym TRY.
Unspecific endonucleases (MWM III DNA): Not detectable in up to 200 μg
after 16 hours incubation at +37°C.
Nicking activity (pBR322 DNA): Not detectable in up to 200 μg after 16 hours
incubation at +37°C.
Ribonucleases (MS2 RNA): Not detectable in up to 40 μg after 16 hours
incubation at +37°C.
DNA (Threshold): ≤10 pg/mg enzyme
Bioburden: ≤125 CFU/g
Stability: At +2 to +8°C within specification range for 18 months.