Silver stain protocol

Andreas M. Stolpe stolpe at MZF.MPIMF-HEIDELBERG.MPG.DE
Wed Jan 24 05:05:31 EST 1996


On Tue, 23 Jan 1996, Marc Visconti wrote:

> Could someone please email me the recipe for silver staining 
> polyacrylamide gels.  We are trying to cut our expenses down and the 
> BioRad reagents are just too expensive.  Thanks in advance.
> 
> --Marc Visconti
> Hospital for Sick Children
> Toronto, Canada
> 
> marcvis at sickkids.on.ca
> 
> 

Hi Marc

We are using the following procedure for silver staining of SDS-PAGE.
This method works very well.

Hope this helps 

Solutions for Silver Stain:

Fixation:	5 % TCA
Reducer:	30 mL ethanol  65 mL H2O 2.5 mL 4 M Na-acetate 400 uL  2
N  Na2H2SO4. adjust to pH 6.0 fill up to 100 mL
Silvernitrate sol.:	0.1 % AgNO3 in H2O  25 uL  formaldehyde (37 % 
Merck) / 100 mL
Developer:	2.5 % (w/v) Na2CO3  50uL  formaldehyde / 100 
mL  (solution should be made directly before use)
Quencher:	1 % (v/v) acetic acid  2% (v/v) of  50 % glycerol


Fix gel in 100 mL TCA for at least 30 min.
incubate in reducer for 30 min.
Wash 3 times with water (3 * 10 min).
Silvernitrate sol. for 30 min.
Briefly rinse the gel in H2O, then 2 changes of developer.
incubate in developer for 1 - 5 min.
Stop the reaction by changing developer with quencher.



______________________________________________________________________________

Dipl. Biol. Andreas M. Stolpe
Max-Planck Institute f. Medical Research  
Department Molecular Cell Research      
Jahnstr. 29	              	      phone: 0049-(0)-6221-486405
				                 -<0>-6221-351286 (priv.)     
                                      fax:       -<0>-6221-486325
D-69120 Heidelberg                    e-mail:  stolpe at mzf.mpimf-heidelberg.mpg.de

______________________________________________________________________________





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