Colloidal Gold total protein stain

Dima Klenchin klenchin at macc.wisc.edu
Wed Nov 1 13:12:54 EST 1995


In article <477q8p$102 at mserv1.dl.ac.uk>,
   LOGAND <logand at msdos.ensam.inra.fr> wrote:

>Can anyone provide or point to me towards a method to stain total protein 
>on western blots (nitrocellulose) after antibody detection of specific 
>antigen using alkaline phosphatase ?

AFTER will not work - there is no way to avoid blocking membrane for ab
detection, which means your total protein is all over the blot. 

BEFORE is easy. Ponceau S is reversible stain (one drawback - not vry 
sensitive). 

Stock: 2% solution in 30% TCA, 30% sulfosalicylic acid (store rt). 

Staining: dilute 1:10 with water, stain for 10-20 min. 
Destaining: water or TBS. Then proceed with blocking for ab staining. 

Hope it helps. 

      __
     / /\         Dima Klenchin
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   / / /\ \       klenchin at macc.wisc.edu
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 / /_/__\ \ \     FAX  (608)262-4570 
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