Proteinase K (lyophilized) / 100 mg

Proteinase K
Proteinase K (lyophilized) / 100 mg
pcs

Proteinase K 

recombinant

Synonym: Endopeptidase K

 

Serine protease with a broad spectrum, intended for the digestion of proteins in biological samples. Applied two chromatographic steps allow for complete removal of  DNA, RNAses and DNAses.

Our recombinant Proteinase K is cloned from fungus Engyodontium album and produced in Pichia pastoris. The enzyme activity is identical to market available products from world leading competitors.

 

 

 Features

 Specifications

 Form

 ultrapure lyophilized powder

 Size

 100 mg  

 Storage

 +4°C

 Activity

 >600mAU/ml; >30 U/mg

 Purity

 - DNA and RNA free
 - DNAse and RNAse free

 Applications

 - purification of target material from contaminating proteins 
 - removal of DNases and RNases when isolating DNA and RNA 
 - Prion (TSE) diagnostics
 - Mitochondria isolation

 
Proteinase K is manufactured and available also in bulk quantities. 
For more information, please contact us info@aabiot.com
 
 

Protocol

MSDS

Leaflet

References: A. Burkiewicz, S. Dąbrowski, P. Barski.  Polish recombinant proteinase K. Postępy Biochemii 2007: 53(4) 327-328.

 
More information...

The enzyme was discovered in 1974 in extracts of the fungus Engyodontium album (formerly Tritirachium album). Proteinase K is able to digest native keratin (hair), hence, the name "Proteinase K". The predominant site of cleavage is the peptide bond adjacent to the carboxyl group of aliphatic and aromatic amino acids with blocked alpha amino groups. It is commonly used for its broad specificity. This enzyme belongs to Peptidase family S8.

 

Endopeptidase activity comparison of commercially available Proteinases K versus A&A Biotechnology Proteinase K

 HPLC Gel-filtration of Proteinase K

Proteinase K Activity

The activity assay was performed over the wide range of enzymes concentrations (mg/ml) at 50°C in 10 mM Tris-HCl pH 8.0 buffer at presence of 2% azo-casein as a substrate. The reaction was terminated by addition of 25% TCA, after 15 min.


Column: BioSep-SEC-s3000 (Phenomenex)
Apparatus: BioCad Sprint (PerSeptive Biosystems)
Dimensions: 500 x 7.8 mm
Mobile Phase: 20 mM Sodium Acetate/Acetic Acid pH 4.5, 1 mM CaCl2
Flow Rate: 0.5 mL/min
Temperature: Ambient
Detection: UV @ 280 nm
Sample volume: 0.1 mL Proteinase K (1 mg/ml) (retention time 48.81 min).
Calculated purity: 99.8%             

 

 Comparison of activities at different temperatures of commercial proteinases K and proteinase K manufactured by A&A Biotechnology    The results of chromosomal DNA isolation by means of A&A Biotechnology DNA extraction kits combined with, either proteinase K from A&A (lanes A) or proteinase K available from competitor R (lanes B)

The assay was performed with 2 μg of each proteinase K at 25, 50 and 70°C, in 10 mM Tris-HCl pH 8.0 buffer at presence of 2% azo-casein as a substrate. The reaction was terminated by addition of 25% TCA, after 15 min.

  Sources od material (isolation kit used): 

  • Tissue (Genomic Mini AX Tissue kit)
  • E.coli / pUC19 plasmid culture (Genomic Mini AX Bacteria kit)
  • Blood (Genomic Mini AX Blood kit)
  • Yeast culture - P.pastoris (Genomic Mini AX Yeast kit)
  • M1 - Lambda phage DNA
  • M2 0 molecular DNA weight marker - Lambda DNA/AvaII (A&A Biotechnology)

 

 

 
 
 

 

             

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