Hipure Proteinase K - China

Cas No: 39450-01-6 E.

C: 3.4.21.64

Cat No: E20041 Specific Activity: 40 U/mg protein

Form: Lyophilized Powder and Liquid Solution are both available

Synonyms: Peptidase K, Endoproteinase K, Endopeptidase K Description

HiPure Proteinase K is an endolytic protease that cleaves peptide bonds at the carboxylic sides of aliphatic, aromatic or hydrophobic amino .

The HiPure Proteinase K is classified as a serine protease.

The smallest peptide to be hydrolyzed by this enzyme is a tetrapeptide.

Features

Active in a wide range of reaction products 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 Storage

Store the HiPure Proteinase K powder at 4ºC or -20℃.

Dissolved Proteinase K solution should be stored at -20ºC.

.

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.

Each lot was tested to ensure the absence of Nucleases and DNA.

Definition of Activity Unit

One unit of the enzyme liberates Folin-positive amino and peptides corresponding to 1 µmol tyrosine in 1 min at 37°C, pH 7.5 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.

Source

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

Molecular Weight

28.9 kDa monomer Dilution Buffer

50 mM Tris-HCl (pH 7.5), containing 5 mM calcium chloride and 50% (v/v) glycerol.

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.

Advantage

The recombinant enzyme is a mutant to the native protease, which gains higher specific activity and yield as well as wider pH and temperature range .

The large scale recombinant preparation has advantage in lot-to-lot consistency, superior purity and cost-efficiency.

DNA-free nature of recombinant Proteinase K made it well-suited in isolating PCR and RT-PCR templates.

Preparation Notes

Activators: 1~5mM Ca2+

To stimulate proteinase K activity, 1-5 mM Ca2+ can be added.

Optimization using activators can increase proteinase activity significantly.

Enzyme activity will be reduced by 25% when calcium is removed by addition of EDTA.

Enzyme activity will be reduced by 80% if the EDTA-Ca2+ complex is removed from the enzyme solution by gel filtration, while it can be partially restored by addition of excess Ca2+.

Inhibitors: DIFP or PMSF

The enzyme is inactivated by DIFP or PMSF (PMSF used at final concentration 5 mM.

).

However, it is not inhibited by EDTA, iodoacetic trypsin-specific inhibitor TLCK, chymotrypsin-specific inhibitor TPCK, and p-chloromercuribenzoate.

Note

• Optimum activity at 50-55°C.

• Rapid denaturation of enzyme occurs at temperatures above 65°C.

• The recommended working concentration for Hipure 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.

• Stable over a wide pH range: 4.0-12.5, optimum pH 7.5-8.0.

Company Contact:

• Posted By: Angen Biotech Co.,Ltd

• Phone: 02038555004

• Address: 8 Yayingshi Road, Luogang District, Guangzhou , Guangdong , China

• Email:

• Website: https://www.angenbio.com

• Business Description:
  
  ANGEN, a leading biotechnology company in China, provides professional biology products and technical service to customers in life science research field.
  
  In 2011, the company was founded by three scientists with a mission to produce innovative and cost-effective products for life science research.
  
  AnGen Biotech's headquarter, R&D, and manufacturing facility are located in Guanghzou.
  
  Now, the comany has more than 10 biologists and 15 sales officer in China.
  
  The comany has about 20 distributors in different cities in local market.
  
  And also have more than 35 customers oversea maily in Noth America, South America, Europe and Asia.
  
  We are targetting to help molecular companies which supplying DNA/RNA prufication reagents.
  
  AnGen Biotech was a professional company in producing Nucleic Purification Accessories like Plasmid DNA Extraction Mini Spin Columns, Genomic DNA Extraction Columns, Gel Purification Columns, RNA Extraction Columns, Maxi Spin Columns.
  
  And also Plastic materies like PCR Tubes, Centrifuge Tubes, Plastic Bottles and Storage Vials.