Staining nitrocellulose

Curt Ashendel ashendel at
Mon Feb 27 09:26:12 EST 1995

On Sun, 26 Feb 1995 23:54:09 -0800, 
Mary Jane Nather  <natherm at> wrote:

>I have been looking for a procedure to stain nitrocellulose membranes 
>after transferring proteins from the polyacrylimide gel.  I have managed to 
>find out that I can use Coomassie, Amido Black, or Ponceau S, but I am 
>unable to determine the advantages to each method.  I don't necessarily 
>care if the stain is reversible, and we are amplifying the protein of 
>interest, so I don't think concentration should be a problem.
>Any insights?  Does anyone have a preferred stain?

No insight, just a preference: Coomassie.  I haven't tried the others. 

Same solution as for staining gels 1% dye in 30% MeOH 10% acetic acid 
(though the solvent concentrations can be varied somewhat).  Immerse 
nitrocellulose for 10 to 15 seconds with agitation then destain for 1 to 
20 min in same solvent lacking dye, until background is acceptable. This 
may be done after blocking and probing, but probing does not work after 
dye staining. This is very fast and very convenient. IT is so convenient 
that I now use it for recording gel results instead of staining, 
destaining, and drying gels (saving the vacuum pump from all that acid).  

Curt Ashendel
Purdue University
West Lafayette, IN
ashendel at

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