Introduction to Proteinase K

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Proteinase K is a subtilisin protease isolated from Candida albicans, CAS number 39450-01-6. It solubilizes histones, degrades miscellaneous proteins, and inactivates lysozyme and ribonuclease to release DNA. The proteinase K produced by our company is a dry powder, which can be stored at low temperature for 2 years at 0~4. After dissolving, it is a colorless and transparent liquid, which can be stored at -20°C for half a year.

The following editor of Desheng will take you to understand the three main advantages of proteinase K.

1. Proteinase K has strong resistance to activity and is active under various reaction conditions. It can be used for DNA extraction in a wide pH range (4-12.5) and higher temperature (50-65). Neither EDTA chelator nor SDS detergent could directly affect proteinase K activity. During extraction of purified protein, EDTA is added to precipitate calcium ions. Although calcium ions can maintain the thermal stability of proteinase K, prevent proteinase K from self-dissolution, and regulate the substrate binding site of proteinase K, it affects the results of the experiment and is not a necessary substance to maintain proteinase K.

2. The enzymatic hydrolysis of proteinase K is fast and will not degrade the target product. When lysing cells, proteinase K digests proteins on the cell surface, inhibiting DNase activity to release the isolated DNA. When detergent is added, proteinase K digests and inactivates RNase in solution, thereby isolating RNA.

3. The cleavage ability of proteinase K is extensive. Proteinase K is a nonspecific endoprotease that cleaves ester and peptide bonds linked to the carboxy-terminus of aliphatic, hydrophobic, and aromatic amino acids.

The concentration of proteinase K affects the structure of cells. Therefore, for the use of proteinase K, Desheng recommends a concentration of 50-100 µg/ml for protein removal and inactivation of enzymes; a concentration below 2mg/ml for tissue processing; 50mM TRIS-HCL buffer and 10mM Concentration of CaCl is used for keratin consumption. After continuous exploration and technological innovation, Desheng produces proteinase K purification technology, mature freeze-drying process, and can provide matching biological buffers for DNA/RNA extraction and separation experiments , stabilize the pH of the solution.