Hello,
I have a problem with western blot, I'm trying to purify one membrane protein expressed in the brain the MW is around 120 KDa. I'm using in vitro model of 293 cells that express this protein. After many steps of solublisation in RIPA buffer, max amplitud of sonication, max centrifugation, quantification of proteins, I boil the samples for 5min, and load in the gel. I'm unable to see this protein by western !!! if you have any idea, please help, thanks
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