silver staining...

[Christian Radauer]

On Sun, 28 Jul 1996, Arioch wrote:

> Date: Sun, 28 JUL 1996 13:01:43 -0500 
> From: Arioch <sai at topaz.microbio.uab.edu>
> Newgroups: bionet.molbio.methds-reagnts
> Subject: silver staining... 
> 
> hi there,
> 	anybody know if there's a way to re-stain a silver stained gel?
> i've already stained and developed my gels, but am still not getting the
> bands as intense as i want them...yes, i know the most obvious thing here
> is the amount loaded on the gel to begin weith and the concentration of
> it...what i want to know is if there's a way to "re-expose"/"over-expose"
> my stain...would longer staining times in my staining solution work?
> currently i stain for 15 min, rinse with water and develop until the bands
> show up...i suppose i could just lengthen the exposure times...any
> ideas???  thx...cheers...
> 

I stain my gels with an alkaline diamine silver solution and develop them 
with acidic formaldehyde solution. If I get too weak bands I wash the gel 
twice with water (10 min. for a 1 mm thick mini-gel) and restain it. The 
silver reduced during the first staining step (even if invisible) 
catalyzes silver reduction during the second one. So take care not to 
overstain the gel. A drawback of this method frequently is higher 
background.

Hope that helps

Christian

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Christian Radauer
Institute of General and Experimental Pathology
University of Vienna

AKH 3Q                      Phone: +43 (1) 40400 5116
Waehringer Guertel 18-20      FAX: +43 (1) 40400 5130
A-1090 Wien
Austria

e-mail: rc150295 at emb1.bcc.univie.ac.at
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