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Proteinase K is an endolytic protease that cleaves peptide bonds at the carboxylic sides of aliphatic, aromatic or hydrophobic amino acids. Proteinase K is classified as a serine protease. The smallest peptide to be hydrolysed is a tetrapeptide.
DNA Extraction lysis reagent for DNA tests & genotyping: The Proteinase K solution is useful when a small amount of biological samples are processed. Add the enzyme solution to DirectPCR reagents at the final concentration of 0.5-1.0 mg/ml (25-50 μl Proteinase K solution per 1 ml DirectPCR reagent). Follow the instruction manuals for DirectPCR Reagent Protocol/Manual for each blood, blood stain, buccal swab, hair root DNA genotyping. Once diluted in DirectPCR reagents, Proteinase K is stable for ~24 hrs.
Other applications: Proteinase K digests native proteins very effectively. It can therefore be used to remove DNases and RNases when isolating DNA and RNA from tissues or cells.
Usage Conditions:
• Proteinase K is active over a wide pH range—optimal activity between 6.5 and 9.5
• Under denaturing conditions—e.g., in the presence of SDS or urea
• In the presence of metal chelating agents—e.g., EDTA
• At high temperatures—optimum digestion temperature is 65°C
Unit Definition: One unit of the enzyme liberates folin-positive amino acids and peptides corresponding to 1μmol tyrosine in 1min at 37°C using denatured hemoglobin as substrate.
Quality Control: Tested for the absence of endo-, exodeoxyribonucleases and ribonucleases
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文献和实验Proteinase K is a serine protease and the main proteolytic enzyme produced by the fungus Tritirachium album Limber (1 ). The enzyme has abroad specificity, cleaving peptide bonds C-terminal to a number of amino acids. The enzyme is produced
RNA Extraction by the Proteinase K Method
. Next the polysomes are dissociated with SDS and proteinase K and finally the protein is removed by several phenol/choloroform extractions (1 ,2 ).
分子量: 28.9 kDa 单位定义: 在250 μl反应体系, 37℃条件下1分钟内能分解释放相当于1 μmol酪氨酸的folin-阳性氨基酸和肽所需要的蛋白酶K的量定义为一个单位(1) 贮存条件: 20 mM Tris-HCl (pH 7.4), 1 mM CaCl2和50%甘油。贮存于-20℃ Proteinase K,中文名为蛋白酶K。不含DNase,不含
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