Huge background with HT22 cells phophoERK antibody - (Dec/12/2011 )

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

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.