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which one is first phosphorilated or total protein?

which one is first phosphorilated or total protein? - Western Blot Forum

which one is first phosphorilated or total protein? - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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Old 07-12-2010, 06:43 AM
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Default 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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