{"product_id":"proteinase-k-powder","title":"Proteinase K Powder","description":"\u003cmeta charset=\"UTF-8\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003eBackground\u003c\/p\u003e\n\u003cp data-mce-fragment=\"1\"\u003eWhat is proteinase K used for? Proteinase K is commonly used for protein digestion, DNA extraction, RNA purification, direct PCR, and genotyping. Since it very effectively inactivates DNases and RNases, the recombinant proteinase K enzyme is added during nucleic acid preparations to isolate highly native, undamaged DNAs or RNAs; it is used in direct PCR kits for genotyping by destruction of proteins in cell lysates (tissue or cell culture cells) and for release of genomic DNA.\u003c\/p\u003e\n\u003cp data-mce-fragment=\"1\"\u003eWhat is the function of proteinase K? Proteinase K is a broad-spectrum serine protease originally isolated from fungus Engyodontium album. The protease was named “Proteinase K” for its ability to digest Keratin. Crystal and molecular structure studies suggest that the enzyme belongs to the subtilisin family characterized with a catalytic triad (Asp39-His69-Ser224) in active site. The proteinase K enzyme has no pronounced cleavage specificity and preferential cleavage site is the peptide bond adjacent hydrophobic amino acids.\u003c\/p\u003e\n\u003cp data-mce-fragment=\"1\"\u003eThe proteinase K enzyme exhibits higher proteolytic activity in the presence of reducing agents such as 5 mM DTT. Proteinase K is inhibited by serine protease inhibitors such as PMSF, DFP, and AEBSF.\u003c\/p\u003e\n\u003cp data-mce-fragment=\"1\"\u003eProteinase K is active in 1% Triton X-100 and fully active in 0.5% (w\/v) SDS which denatures protein substrates to increase digestion rates. The enzyme works best at 50-200 ug\/mL at pH 7.5-8.0, 37 °C and is usually denatured by subsequent phenol extractions. Incubation times vary from 30 minutes to 18 hours and proteinase K can auto-digest during long incubation.\u003c\/p\u003e\n\u003ch3 data-mce-fragment=\"1\"\u003eProteinase K Powder\u003c\/h3\u003e\n\u003cp data-mce-fragment=\"1\"\u003eThe recombinant proteinase K enzyme is from yeast cells with cloned gene encoding Engyodontium album (Tritirachium album) endolytic protease. The recombinant proteinase K 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.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eCAS No.\u003c\/strong\u003e: 39450-01-6.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eE.C.\u003c\/strong\u003e: 3.4.21.64.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eMolecular Weight\u003c\/strong\u003e: 29.3 kDa.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003epI\u003c\/strong\u003e: 8.9.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003epH range\u003c\/strong\u003e: 4.5-12.0; high activity in the pH range of 7.5-11.5.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eUnit Definition\u003c\/strong\u003e: One unit of the proteinase K enzyme is defined as the enzyme activity that produce 1 umol of tyrosine per minute from casein at 37°C at pH 7.5.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eSpecific activity\u003c\/strong\u003e: ≥ 34 units\/mg of protein.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eTemperature profile\u003c\/strong\u003e:  37-70°C temperature range recommended; maximum activity at 70°C.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eExtinction coefficient\u003c\/strong\u003e:  E\u003csup data-mce-fragment=\"1\"\u003e1%\u003c\/sup\u003e = 14.2; 280 nm, 10 mM NaCl and 5 mM CaCl2, pH 8.0.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003ePurity\u003c\/strong\u003e: \u0026gt;99.5% by reducing SDS-PAGE. Each lot of the proteinase K (powder and solution) was tested to ensure the absence of Nucleases and DNA. DNase is not detected in quality control procedure of incubation 40 µg Proteinase K with 1 ug λ DNA for 6 hours at 37°C. RNase is not detected in quality control procedure of incubation 40 µg Proteinase K with 2 ug RNA for 2 hours at 37°C. The preparation is considered as RNase and DNase free.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eFormulation\u003c\/strong\u003e: lyophilized powder.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eReconstitution\u003c\/strong\u003e: The proteinase K enzyme is soluble in water (1 mg\/mL), yielding a clear colorless solution.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eSolution preparation\u003c\/strong\u003e: The molecular biology grade recombinant proteinase K, lyophilized powder is NOT sterile. It can be used to make molecular biology grade proteinase K solution, PCR grade proteinase K solution, NGS grade proteinase K solution, CHI grade proteinase K solution, and so on. The proteinase K stock solution can be prepared as a 20-40 mg\/ml in 20 mM Tris-HCl buffer, sterilized using a 0.22 µm filter and supplied in 50% sterilized glycerol to store in aliquots at wide temperature range from 24°C to -80°C. PES or PVDF membranes with low protein binding are recommended in sterile filtration. We offer irradiation sterilization option to bulk quantity order. The Gamma irradiation procedure may cause slight enzyme activity loss. Please inquiry.\u003cbr data-mce-fragment=\"1\"\u003eStorage buffer: 20 mM Tris-HCl, 1 mM CaCl2, 50% Glycerol, pH 7.4.\u003cbr data-mce-fragment=\"1\"\u003eDilution buffer recommended: 20 mM Tris-HCl (pH 7.4), 1 mM CaCl2; or 20 mM Tris-HCl (pH 7.4), 1 mM CaCl2, and 2% Glycerol.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eActivators\u003c\/strong\u003e: 1-5 mM Ca\u003csup data-mce-fragment=\"1\"\u003e2+\u003c\/sup\u003e. To stimulate proteinase K activity, 1-5 mM Ca\u003csup data-mce-fragment=\"1\"\u003e2+\u003c\/sup\u003e can be added. Optimization using activators can significantly increase proteinase activity. 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-Ca\u003csup data-mce-fragment=\"1\"\u003e2+\u003c\/sup\u003e complex is removed from the enzyme solution by gel filtration, while it can be partially restored by addition of excess Ca\u003csup data-mce-fragment=\"1\"\u003e2+\u003c\/sup\u003e.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eInhibitors\u003c\/strong\u003e: DIFP or PMSF. The proteinase K enzyme is inactivated by DIFP or PMSF (PMSF at the final concentration of 5 mM) but not by EDTA , iodoacetic acid, trypsin-specific inhibitor TLCK, chymotrypsin-specific inhibitor TPCK, and p-chloromercuribenzoate.\u003c\/p\u003e\n\u003cp data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eShipping\u003c\/strong\u003e: The recombinant proteinase K powder is shipped with ice packs. Upon receipt, store it immediately at the temperature recommended below.\u003cbr data-mce-fragment=\"1\"\u003e\u003cstrong data-mce-fragment=\"1\"\u003eStability \u0026amp; Storage\u003c\/strong\u003e:\u003cbr data-mce-fragment=\"1\"\u003eUse a manual defrost freezer and avoid repeated freeze-thaw cycles.\u003cbr data-mce-fragment=\"1\"\u003e\u0026gt;2 years from date of receipt if stored at -20 to -70°C.\u003cbr data-mce-fragment=\"1\"\u003e\u0026gt;1 years from date of receipt if stored at room temperature.\u003cbr data-mce-fragment=\"1\"\u003e\u0026gt;1 years from date of receipt if stored at 2–8°C.\u003c\/p\u003e","brand":"Apenzy Biosciences","offers":[{"title":"1 g","offer_id":47556835213591,"sku":"MB111-01","price":175.0,"currency_code":"USD","in_stock":true},{"title":"10 g","offer_id":47558218252567,"sku":"MB111-10","price":1400.0,"currency_code":"USD","in_stock":true},{"title":"25 g","offer_id":47558218285335,"sku":"MB111-25","price":2800.0,"currency_code":"USD","in_stock":true}],"url":"https:\/\/apenzybio.com\/products\/proteinase-k-powder","provider":"Apenzy Biosciences","version":"1.0","type":"link"}