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Applications : Antigen, Drug related studies, Protein structure analysis (Crystal / electron microscope)
Review: Used proteinase K to improve the purity of the extracted DNA from a Bacillus thermoamylovorus strain producing a thermotolerant lipase by digesting contaminating cell proteins. To check the effectiveness of proteinase K to inhibit the activity of purified lipase in the absence and presence of PMSF.
By Shamsher S Kanwar
What is the activity of Proteinase K in common buffers?
What is the recommended pH for Proteinase K to keep its enzyme activity?
What is the main applications of Proteinase K?
How to dissolve proteinase K powder? What should I do if the dissolution is not complete or the dissolution time is longer ?
What is the long-term storage temperature of proteinase K from CUSABIO?
Whether the form of proteinase K is liquid or powder, will repeated freezing and thawing affect the activity?
What is the recommended working concentration of Proteinase K from CUSABIO?
What is the optimal temperature for proteinase K activation? And how to inactivate proteinase K?
Proteinase K digest protein and remove contamination, but it doesn’t digest itself. Why?
How long do the samples need incubating with Proteinase K?
Does EDTA inactivate proteinase K?
How does Proteinase K works in DNA extraction?
Can proteinase K degrade DNA?
Instead of degrading the DNA, protease K digests the proteins surrounding the DNA, allowing it to be isolated intact.
What does proteinase K do to the cell?
Proteinase K can lyse or break down the cell membrane and release the intercellular materials including DNA and RNA.
How do you use proteinase K?
Dissolve lyophilized proteinase K in deionized sterile water. We recommend to add 10 mM Tris HCl, pH 7.5, 20 mM calcium chloride, and 50% glycerol, and then aliquot for long-term storage at -20/-80°C.
Why do we denature proteinase K?
Because proteinase K may degrade the useful proteins in the downstream applications of isolated nucleic acids.
How do you make proteinase K buffer?
The best buffer for proteinase K will be different in various applications. Proteinase K is active over a wide pH range (4-12.5) and relatively high temperature (optimum temperature 50-65 ˚C). Generally, proteinase K is stable and keeps highly active in buffers that contain denaturing reagents such as urea, sodium dodecyl sulfate (SDS), and guanidinium salts.
Why do we add proteinase K to our DNA extractions?
Proteinase K is used to lyse cells, disrupt tissue integrity, and degrade various proteins to make the pure DNA is fully released during DNA extraction. It also inactivates DNase present in the sample, ensuring DNA whole and intact. The wide temperature range and relatively strong pH tolerance of proteinase K also contribute to its extensive availability in nucleic acid extraction.