Anyone knows if it's ok to prepare RIPA buffer without EGTA?
My intention is to use this RIPA buffer to lysate cells for a western blot of b-catenin and phospho b-catenin. I have everything it's necessary to prepare the buffer (including NaCl, Tris, EDTAm, SDS, Triton, Sodium deoxycholate and protease and phosphatase inhibitors) except EGTA...
I appreciate any help or advice.
Thank you.
EGTA and EDTA adding both is better, but I think you know the answer " You dont have EGTA", then go ahead with the EDTA. Both EDTA, and EGTA have similar properties (EGTA have higher affinity for ca). As long you have all protease inhibitors, and EDTA, I dont think that skipping EGTA will cause any problem for your prep. I hope your protein dont have stability issue because of metal chelation.
Hi there,
The issue is actually the possible effect of the absence of EGTA on your protein of interest during extraction... Have a go and see!
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