Experimentally improved reliability of ultrasensitive silver staining of protein in polyacrylamide gels

✍ Scribed by Margret Eschenbruch; Robert R. Bürk

• Publisher: Elsevier Science
• Year: 1982
• Tongue: English
• Weight: 785 KB
• Volume: 125
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(82)90387-6

✦ Synopsis

Variations of the ultrasensitive silver staining method of B. R. Oakley, D. R. Kirsch, and N. R. Morris (Anal. Biochem. 105, 361-363 (1980)) have been tested. It was established that the reliability of the method was greatly improved if (i) free silver was carefully washed out before reduction with formaldehyde; (ii) the extent of development was controlled by using methylamine to inactivate the formaldehyde; and (iii) the optimum quantity of ammonia, which was found to be 4 mol/mol of silver was used (this quantity was defined as that which titrates a particular amount of 1 N HCl). The time of preparation of the formaldehyde reducer was found not to be highly critical. In our hands the method can detect down to 0.1 ng of protein/S-mm slot.