[Protein-analysis] Re: Doing a western after coomassie staining

Dr Engelbert Buxbaum via proteins%40net.bio.net (by engelbert_buxbaum from hotmail.com)
Tue Feb 20 09:20:19 EST 2007


Coomer wrote:

> Does anyone know a protocol to transfer protein after staining the gel
> with coomassie (10% Acetic acid and 50% EtOH, 0.2% Brilliant Blue) ?

Can be done with Dunns blotting buffer with 50 uM SDS added (or Towbins
buffer without methanol). Preincubate the gel with the buffer for 15-30
min to allow SDS-binding to the proteins. Works even if the gels had
been dryed, if they are re-hydrated in the buffer.

Note however that the yield will be lower than with unstained gels.


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