Proteinase K (also known as endopeptidase K) is an extracellular endopeptidase isolated from a culture filtrate of the fungus Tritirachium album limber. This unique fungus can degrade keratin (hence Proteinase “K”) and can grow using the by-product as its sole source of carbon and nitrogen.
The enzyme is stable over a broad temperature and pH range, with optimum activity at 55-65°C and pH 8.0. It’s also stable in high salt concentrations and active in the presence of strong detergents such as SDS. It has a molecular weight of 28.9 kDa and it shows fairly low specificity for its polypeptide targets.
Features and Benefits of AG’s Proteinase K
• DNAse and RNAse free via chromatography purification
• 6x greater activity than Pronase and 3x greater activity than bovine trypsin
• Broad substrate specificity
• Versatile in blood, cultured cells, tissues, bacteria, yeast, etc.
• Active and stable over a wide range of conditions
• Compatible with most DNA and RNA isolation buffers
Applications in Next Generation Sequencing (NGS) and Microarray Technologies
• Nucleic acid purification by inactivating nucleases when extracting DNA and RNA from yeast, bacteria, and mammalian cell & plant cell lysates
• Improves cloning efficiency of PCR products
• Sample preparation for quantification of DNA adduct levels by accelerator mass spectrometry
• Inactivation of enzyme cocktails in ribonuclease protection assays
• Added to extraction procedures to optimize RNA yields from primary breast tumors for microarray studies
Applications with Proteins
• Detection of Bovine Spongiform Encephalopathy proteins which are uniquely resistant to proteolytic degradation.
• Tissue digestion as an alternative sample preparation approach for quantitative analysis using liquid chromatography—tandem mass spectrometry
• Specifically modify cell surface proteins to analyze membrane structures for protein localization
• Generates protein fragments used in characterization of functional studies
|Synonyms||Endopeptidase K, Protease K|
|Appearance||White to off-white lyophilized solid|
|Solubility||50mM Tris-HCl, 2mM calcium acetate, pH 8.0|
|Storage Temp||Store desiccated at +4°C. Store in tightly sealed vial. WARNING: HARMFUL! IRRITANT!|
|Use||Proteinase K is a stable and highly reactive serine protease|
|Handling||Store desiccated at +4°C. Store in tightly sealed vial.|
|msds 1||P-1265, Proteinase K, SDS, diamond format.pdf|
|Certificate of Analysis 1||P-1265, J1298.pdf|
|Certificate of Analysis 2||P-1265, J1072I.pdf|
|Certificate of Analysis 3||P-1265, J1072H.pdf|
|What is proteinase K used for?||Proteinase K is commonly used in molecular biology to digest protein and remove contamination from preparations of nucleic acid.|
|What is the quickest most effective way to inactivate proteinase K?||As with most protein enzymes, change the temperature or change the pH significantly.|