Western Blot staining

Dennis J. Templeton djt2 at po.CWRU.Edu
Thu Nov 12 18:06:34 EST 1992


In a previous article, sandy at pt.Cyanamid.COM (Sandy Silverman) says:

>We once read about a method to detect protein on a Western blot prior
>to antibody detection.  It involved a red dye and acetic acid.  It was 
>reversible.  Does anyone have the reference?   Thanks-----Sandy
>
We use a recipe that is simpler than the Sambrook one:

0.5% Ponseau Red
1% Acetic acid
in water

We stain Immobilon filters for ~1 min and then rinse under the DI water tap 
for about 1 minute.  The limit of sensitivity is about 200-400 ng, or 
maybe worse.

On immobilon, you can detect proteins even more sensitively by simple
observation; there is a shiny-ness that is evident at the right angle. 
Personally, I mark my lanes and marker proteins this way, but others in my
lab can't see it and prefer the ponseau stain.

good luck,

dennis



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