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Huge background with HT22 cells phophoERK antibody

Western blotting he

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#1 sikander

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Posted 12 December 2011 - 03:18 AM

Hi all,

I am studying the ERK activation in HT22 cells and for this first i probe my blot with total ERK and got good signal and later on when i do stripping. After stripping the blot i prob the blot with phosphoERK antibody.

My total ERK gives very good signal, while there comes huge background in the phospho ERK probing.

Can anyone gives some suggestion.

thanks

#2 Biouday

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Posted 12 December 2011 - 04:40 AM

Hi

Did you check blots only with phospho ERK, is that good without background.
And my suggestion is always do PhoshoERK ( any phospho protein) before doing the total protein.




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