www.bioland-sci.com

Proteinase K (Lyophilisate, 1g)

Price: $549.00

Proteinase K is an endolytic protease that cleaves peptide bonds at the carboxylic sides of aliphatic, aromatic or hydrophobic amino acids. The Proteinase K is classified as a serine protease. The smallest peptide to be hydrolyzed by this enzyme is a tetrapeptide.
 
Specific Activity: 40 U/mg protein
Molecular Weight: 28.9 kDa monomer
Source: Pichia pastoris cells with a cloned gene encoding Tritirachium album endolytic protease (Proteinase K).
 
Applications
  • Isolation of genomic DNA from cultured cells and tissues
  • Removal of DNases and RNases when isolating DNA and RNA from tissues or cell lines 
  • Determination of enzyme localization
  • Improving cloning efficiency of PCR products 
Quality Control
  • DNase Activity: None detectable enzyme activity with λ DNA after 6 hrs incubation at 37ºC.
  • RNase Activity: None detectable ribonuclease activity after 16 hrs incubation at 25ºC. 
Dilution Buffer
50 mM Tris-HCl (pH 7.5), containing 5 mM calcium chloride and 50% (v/v) glycerol. 

Definition of Activity Unit
One unit of the enzyme liberates Folin-positive amino acids and peptides corresponding to 1 µmol tyrosine in 1 min at 37°C using denatured hemoglobin as substrate.
Enzyme activity is assayed in the following mixture: 0.08 M potassium phosphate (pH 7.5), 5 M urea, 4 mM NaCl, 3 mM CaCl2 and 16.7 mg/ml hemoglobin.

Storage 
For long time storage, store the Proteinase K powder at 4ºC and solution at -20℃. 

Inhibition and Inactivation
  • Inhibitors: Proteinase K is not inactivated by metal chelators, by thiol-reactive reagents or by specific trypsin and chymotrypsin inhibitors. Phenylmethylsulfonyl fluoride and diisopropyl phosphorofluoridate completely inhibit the enzyme.
  • Inactivated by heating at 95°C for 10 minutes. 
Other Note:
  • Optimum activity at 50-55°C. 
  • Rapid denaturation of enzyme occurs at temperatures above 65°C. 
  • The recommended working concentration for Proteinase K is 0.05-1 mg/ml. The activity of the enzyme is stimulated by 0.2-1% SDS or by 1-4 M urea.
  • Ca2+ protects Proteinase K against autolysis, increases the thermal stability and has a regulatory function for the substrate binding site of Proteinase K.
  • Stable over a wide pH range: 4.0-12.5, optimum pH 7.5-8.0.

Add to Cart:

  • Model:
    [EPD02-02]
  • Shipping Weight: 1lbs

Copyright © 2018 Bioland Scientific, LLC. All rights reserved. Powered by Zen Cart. Hosted by Siteground.
FedEx service marks used by permission.