Hi
I want to know the protocol of using M-per lysis buffer for protein extraction.
I have came across many protocols which says to use it alone.
Some says use it in combination with Protease inhibitor and few others recommend to use PhosSTOP(phosphatase inhibitor) + Protease inhibitor cocktail + M-per.
As per my understanding, PhosSTOP maybe used when we may want to prevent dephosphorylation.
But I am confused of the fact that whether protease inhibitor cocktail is necessary to be added or not.
Anyone who uses M-per, kindly may clear my doubt.
Thanks in advance!
M-PER is nondenaturing, meaning that any proteases in the extract could still be active. The user guide for M-per says that protease inhibitors may be added to the reagent. If you are concerned about proteolysis in your lysate, it would be a good idea to add a protease inhibitor cocktail.
Agreed! I think as a general rule, always add protease inhibitors to any lysis buffer that does not contain them as components! If you plan to look for phosphorylations you will also need to add the phosphatase inhibitors.
Thank you Eugènia Almacellas . I am adding phosphatase inhibitor with protease inhibitor as well.
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