I've operated left side and collected left part of the brain and I've been used RIPA buffer for homogenization and centrifuged at 10,000 rpm for 20 min,the occlusion period is 60 min.all my 24hr samples(whole cell lysate) are showing significant decrease in P-JNK expression,which is contradictory to available data .Is this technical problem or natural .If anyone had encountered same problem please help me out.
Hello Sireesh,
I think your experimental conditions are okay. Could you re-check with the previous result's protocols, I mean experimental conditions, drug doses, types of instruments used? Because, these also contributed greatly to your experimental outcomes as you are looking for the phosphorylated proteins.
Dear moniruzzaman,
Thanks a lot for your suggestion,i can crosscheck one more time with all those parameters.
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