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phosphorylated protein Western blotting - HELP!! (Jul/13/2006 )

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Hi bonnie,

I'm working with phosho-antibodies in colon tissues. In my lysis buffer (midified RIPA buffer) there are some phosphatase inhibitors such as okadaic acid and beta-glycerophosphate and other protease inhibitors. I wonder that did your phospho-antibody work in tissues at the end of your effort? Because I have not seen any bands with colon tissues. Thanks.

QUOTE (bonnie @ Jul 25 2006, 09:15 AM)
QUOTE (Missele @ Jul 14 2006, 07:56 AM)
what do you mean by ANY signal?
you can't see proteins, or you can't see phosphorylated proteins?

i check the protein transfer by ponchu.
And transfer was well done.

but i i couldn't detect the phosphorylated protein on film after ECL detection.


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