Analytical Biochemistry 1996-12-15

Negative staining of proteins in polyacrylamide gels with methyl trichloroacetate.

G Candiano, M Porotto, M Lanciotti, G M Ghiggeri

Index: Anal. Biochem. 243(2) , 245-8, (1996)

Full Text: HTML

Abstract

This paper describes a new, sensitive (in the nanogram range), and rapid (two-step) technique for the negative staining of proteins in polyacrylamide gels in the presence or absence of sodium dodecyl sulfate. After separation, gels are incubated with 8% methyl trichloroacetate ester in 38% isopropanol and then washed in water to produce a negative image of colorless proteins against an opaque background. The technique allows unmodified proteins to be recovered for biological studies or transblot for amino acid sequence. Finally, owing to the reversibility of the process, gels can be restained after rapid visualization. For these reasons, negative staining with methyl trichloroacetate should become the method of choice for rapid and sensitive staining of proteins prior to further processing, including stable staining with silver ions.