western blot help!!!

karthij96 at yahoo.com karthij96 at yahoo.com
Sat Apr 19 11:59:56 EST 2003


i have just started doing westerns recently.  i am facing problems with the PVDF membrane that 
we use.  i soak it in methanol  & before it dries i soak it in ddwater & then in the western transfer 
buffer.  
The transfer looks good--as i am able to see all the lanes & even the marker .  The problem is 
that after i block the membrane in 5% milk ( carnation non-fat dry milk powder &  made in 1X 
TBS) for an hour or even overnite in a cold room---the membrane refuses to wet---it looks as if 
its dry, only the egdes seem to be wet.
 Anyone with a solution---will be greatly appreciated.
thank you
aditi

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