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-   -   which one is first phosphorilated or total protein? (http://www.molecularstation.com/forum/western-blot-forum/73343-one-first-phosphorilated-total-protein.html)

Pokoke 07-12-2010 06:43 AM

which one is first phosphorilated or total protein?
 
Dear all,

I have transferred my protein in PVDF membrane and I want to determine the expression of phosphorilated EGFR and total EGFR. Which one should I blot first? previously I have blot actin, and then phosphorilated EGFR and then EGFR. I have good result in actin but nothing in both p-EGFR and EGFR. Do the order of blotting influence the result?

Thank you for any advice

SiJohn


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