Previsible silver staining of protein in electrophoresis gels with mass spectrometry compatibility.
A convenient silver staining method for protein in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels is described. The method is previsible, sensitive, and mass spectrometry (MS) compatible. Two visible counter ion dyes, ethyl violet (EV) and zincon (ZC), were used in the first staining solution with a detection limit of 2 to 8 ng/band in approximately 1h. The dye-stained gel can be further stained by silver staining, which is based on acidic silver staining employing ZC with sodium thiosulfate as silver ion sensitizers. Especially, ZC has silver ion reducing power by cleavage of the diazo bond of the dye during silver reduction. The second silver staining can be completed in approximately 1h with a detection limit of 0.2 ng/band.
Jin
Li-Tai
LT
School of Pharmacy, Wenzhou Medical College, Wenzhou 325-000, China.
Li
Xiao-Kun
XK
Cong
Wei-Tao
WT
Hwang
Sun-Young
SY
Choi
Jung-Kap
JK
D053000:Analytic Sample Preparation Methods / Q000191:economics / Q000379:methods*; D000818:Animals; D002417:Cattle; D004396:Coloring Agents / Q000737:chemistry; D003365:Costs and Cost Analysis; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D016014:Linear Models; D013058:Mass Spectrometry; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics; D012394:Rosaniline Dyes / Q000737:chemistry; D012680:Sensitivity and Specificity; D016622:Silver Staining / Q000191:economics / Q000379:methods*; D013997:Time Factors; D015032:Zinc / Q000737:chemistry
D004396:Coloring Agents; D011506:Proteins; D012394:Rosaniline Dyes; C034887:ethyl violet; D015032:Zinc