transfer proteins from a stained gel

HYDE at horta HYDE at horta
Fri Oct 1 10:26:17 EST 1993


In article <01H3K46TKKFW8ZF3FO at minna.acc.iit.edu>, CMSMUELLER at MINNA.ACC.IIT.EDU writes:
>Help!!  Does anyone know how to transfer protein from 
>a gel to nitrocellulose after it has  been stained with 
>Coomassie blue?  We need to perform a Western
>analysis but do not have any more of the sample.  
>
>Thanks
>David Mueller
>Chicago Medical School

Try destaining the gel with your methanol/acetic acid solution that you used 
to remove excess stain. Make sure that the gel is rehydrated as the methanol
tends to dry the acrylamide matrix and make it brittle. After this you may 
transfer to nitrocellulose using standard techniques.

I have had success directly transferring stained gels to nitrocellulose, the
stain will simply be washed out when you begine your probing procedures.

Fred Hyde
3M Company
St Paul, MN



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