Negative staining of proteins in polyacrylamide gels with methyl trichloroacetate.
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.
Candiano
G
G
Department of Nephrology, G. Gaslini Children Hospital, Largo G. Gaslini 5, Genoa, 16148, Italy.
Porotto
M
M
Lanciotti
M
M
Ghiggeri
G M
GM
D011232:Chemical Precipitation; D062845:Chloroacetates; D004396:Coloring Agents; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000737:chemistry; D006801:Humans; D008970:Molecular Weight; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000592:standards; D012015:Reference Standards; D012680:Sensitivity and Specificity; D012867:Skin / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods* / Q000706:statistics & numerical data
D062845:Chloroacetates; D004396:Coloring Agents; D011506:Proteins; C103957:methyl trichloroacetate