澳门赌搏网站大全-澳门网络娱乐场网站大全

Products > DNA Purification Accessary Reagents > Proteinase K (Lyophilized Powder)(N9016)

Proteinase K (Lyophilized Powder)(N9016)

Proteinase K

Catalog #

Size

Price

N9016

100mg


Contents:   Proteinase K     100mg
                  

Specific Activity: 40 U/mg protein
Form: Contains 100 mg Proteinase K, Lyophilized Powder

Features
Active in a wide range of reaction products

Description
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.

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 37?C.
RNase Activity: None detectable ribonuclease activity after 16 hrs incubation at 25?C. 

Source
Pichia pastoris cells with a cloned gene encoding Tritirachium album endolytic protease (Proteinase K).

Molecular Weight
28.9 kDa monomer (6).

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 ?mol 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 4?C and Delution Buffer at -20℃ separately. For use, dissolved solution should be stored at -20?C.

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. 

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.


XML 地图 | Sitemap 地图