Negative staining of proteins in polyacrylamide gels with methyl trichloroacetate

Analytical Biochemistry
G CandianoG M Ghiggeri

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.

Citations

Aug 16, 2005·American Journal of Respiratory and Critical Care Medicine·Giovanni CandianoOlga Zegarra-Moran
Nov 29, 2005·Clinical Chemistry and Laboratory Medicine : CCLM·Isabella LevreriGiovanni Melioli
Apr 25, 2007·Current Eye Research·Silvia RaveraAlessandro Morelli
Jul 2, 2003·Proteomics·Maurizio BruschiPier Giorgio Righetti
Jan 12, 2008·Electrophoresis·Giovanni CandianoGian M Ghiggeri
Sep 29, 2006·Journal of the American Society of Nephrology : JASN·Giovanni CandianoGian Marco Ghiggeri
Jan 12, 2007·American Journal of Physiology. Lung Cellular and Molecular Physiology·Giovanni CandianoOlga Zegarra-Moran
Sep 12, 2019·Scientific Reports·Laura SantucciGiovanni Candiano
Mar 19, 2011·Molecular BioSystems·Laura SantucciGian Marco Ghiggeri
Nov 1, 2003·Expert Review of Neurotherapeutics·Craig Watson
Aug 9, 2005·Journal of Proteome Research·Maurizio BruschiGiovanni Candiano

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