westerns with coomassie stained gels?

Wolfgang Schechinger hubahopp at gmx.de
Thu Nov 4 10:56:54 EST 2004


Thanks, Stefan!

Unfortunately, this paper is not available online yet. Hoewever, I have
come across another one in the same journal:

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_ui
ds=8742090&dopt=Abstract

Meanwhile, a 2hr transfer at standard conditions just seems having
transferred the coomasie as the coloured potein standard is still in gel
but the coomassie staining pattern is on the nitrocellulose.... Will
transfer again over night now.

Best regards,

Wolfgang


At 15:51 04.11.2004 +0100, Stefan Lienenklaus wrote:
>Hi Wolfgang..
>
>Maybe this link answers your question.. - There ist a procedure how to 
>treat Coomassie gels before blotting in part 1.6 of the paper..
>(at least it's said to be) - I don't have the paper myself.. - so I 
>can't tell any details.
>
>Stefan
>
>__________
>
>Anal Biochem. 1986 Jan;152(1):94-9.
>
>
>Protein transfer from fixed, stained, and dried polyacrylamide gels and 
>immunoblot with protein A-gold.
>
>Perides G, Plagens U, Traub P.
>
>The method of electrophoretically transferring proteins from fixed and 
>stained polyacrylamide gels onto nitrocellulose paper has been 
>reevaluated. It is shown that the tedious destaining of gels is not 
>necessary because Coomassie brilliant blue, although it binds 
>tenaciously to nitrocellulose paper, does not reduce the transfer 
>efficiency of proteins. However, its presence impairs the visibility of 
>proteins as detected, for instance, by the immunogold technique. 
>Therefore, a rapid method for the complete removal of the stain from the 
>nitrocellulose paper after completion of the immunogold procedure was 
>developed. Furthermore, it is shown that proteins from dried 
>polyacrylamide gels can still be transferred onto nitrocellulose sheets 
>with an efficiency of approximately 50% compared to proteins transferred 
>from fixed gels.
>
>PMID: 2420231 [PubMed - indexed for MEDLINE]
>
>
Wolfgang Schechinger, PhD (Dr. rer. nat.)
Chemistry and Life Sciences

BGFA XU44
Med Clinic I / Endocrine Research 
University Hospital "Bergmannsheil"
Bürkle-de-la-Camp-Platz 1
44789 Bochum
Germany

Phone +49-234-302-4789
Fax   +49-234-307-4790

wolfgang.schechinger at gmx.net




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