Polyacrylamide gels, silver-stained

As compared to ethidium bromide-stained agarose gels, many more amplified bands can be visualized by separating die products of the reaction on a polyacrylamide gel and staining with silver.9 This gel system also permits detection of bands amplified weakly by primers as short as 5 nucleotides. 

Polyacrylamide, precast and homemade gels Silver staining Rehhein et al. (1999)... 

The protein complexes of P. lutherii were separated from thylakoid membranes on a digitonin (0.1%), sucrose gradient (10-40%). Fractions were collected and the polypeptides separated under denaturing conditions on a 15-20% gradient polyacrylamide gel and stained with Silver stain. Polypeptides from other gels were transferred electrophoretically to nitrocellulose membrane and reacted with antisera to the acf complex of P. lutherii. 

Amido black is a commonly used stain, but it is not very sensitive. It is often used to visualize concentrated proteins or components that are readily accessible to dyes such as proteins that have been transferred from a gel to nitrocellulose paper. Two of the more sensitive and more frequently used stains are Coomassie Brilliant Blue (R250 and G250) and silver stains. Because these stains interact differently with a variety of protein molecules, optimization of the fixative and staining solutions is necessary. The Coomassie stains are approximately five times more sensitive than amido black and are appropriate for both agarose and polyacrylamide gels. The silver stain is approximately 100 times more sensitive than Coomassie and is typically used for polyacrylamide gels. 

Shevchenko, A., Wilm, M., Vorm, O., and Mann, M. (1996). Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal. Chem. 68, 850—858. 

Figure 3 Biosynthesis and purification of 90-kD elastin analogue analyzed by denaturing polyacrylamide gel electrophoresis (10-15% gradient, visualized by silver staining). Lanes 1-7 time course of target protein expression at 0, 30, 60, 90, 120, 150, and 180 minutes after induction. Lane 9 soluble lysate of induced E. coli expression strain BLR(DE3)pRAMl. Lanes 10-13 protein fractions obtained from immobilized metal affinity chromatography of the lysate on nickel-NTA agarose (imidazole gradient elution). Lanes 8,14 protein molecular weight standards of 50, 75, 100, and 150 kD.

Figure 10.7 Polypeptide patterns of material associated with lipid globules of cow s milk. The sodium dodecyl sulfate-polyacrylamide gel in (a) was stained with coomassie blue, and polypeptides in the gel in (b) were visualized with the more sensitive silver stain (Merril ef at. 1981). In both (a) and (b) the left lane contains proteins extracted directly from washed milk...

Merril, C. R., Dunau, M. L. and Goldman, D. 1981. A rapid sensitive silver stain for polypeptides in polyacrylamide gels. Anal Biochem. 110, 201-207. 

A silver-stained gel obtained by electrophoresis of a standard protein mixture under denaturing conditions. Samples were run on 12% polyacrylamide gels,... 

Support Protocol 2 Silver Staining of Polyacrylamide Gels B3.1.15... 

Heukeshoven, J. and Dernick, R. 1985. Simplified method for silver staining of proteins in polyacrylamide gels and the mechanism of silver staining. Electrophoresis 6 103-112. 

Wise, G. E. and Lin, F. (1991) Transfer of silver-stained proteins from polyacrylamide gels to polyvinylidene difluoride membranes. J. Biochem. Biophys. Methods 22, 223-231. 

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