Greetings.
I founded protease K solution, but have some qusetion...
1. what's different between Protease K solution and Protease K buffer? (some solution enclude CaCl2 and no EDTA, or enclude EDTA, or enclude EDTA and SDS...)
2. For solution,
- 20mg/ml proteinase K
- 10mM Tris-HCl(pH 7.5)
- 20mM CaCl2
- 50% glycerol
then, for example, is that OKay I want to make that solution 10ml, mix protease K 200mg and Tris, CaCl2, glycerol 5ml and water pours to 10ml? and, want to make stock solution except PK, for example stock solution is 10X, 100mM Tris-HCl, 200mM CaCl2 and 50% glycerol(v/v)?
https://www.qiagen.com/cn/resources/faq?id=64c67eac-3938-4fa0-8910-4e29e4d0f8f9&lang=en
Proteinase K is a subtilisin-type protease, which cleaves at the carboxyl side of hydrophobic, aliphatic and aromatic amino acids. It is particularly suitable for short digestion times. It possesses a high specific activity over a wide range of temperatures and pH values with substantially increased activity at higher temperature. Soluble calcium is not essential for enzymatic activity. This means that EDTA, which is used to inhibit Mg2+-dependent enzymes such as nucleases, will not inhibit Proteinase K activity. QIAGEN Protease is a broad-specificity Serine protease with high activity, cleaving preferentially at neutral and acidic residues. It is an economical alternative to Proteinase K for isolation of native DNA and RNA from a variety of samples
In addition, proteinase k is soluble in water (1mg/ml)
https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma-Aldrich/Datasheet/p2308dat.pdf
Ozkan Gelincik Thank you for your answer! I will make solute powder proteinase K in solution.
Some solution enclude CaCl2(only, except EDTA), or some solution enclude EDTA(EDTA only or with SDS), both CaCl2 and EDTA don't affect for proteinase K storage?
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