title: Counterion dye staining of proteins in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and tryptic gel digestion of stained protein for mass spectrometry.
abstract: A fast and matrix-assisted laser desorption/ionization mass spectrometry compatible protein staining method in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis is described. It is based on the counterion dye staining method that employs oppositely charged two dyes, zincon and ethyl violet, to form an ion-pair complex. The protocol, including fixing, staining, and quick washing steps, can be completed in 1-1.5 h, depending upon gel thickness. It has the sensitivity comparable to the colloidal Coomassie Brilliant Blue G stain using phosphoric acid as a component of staining solution (4-8 ng). The counterion dye stain does not induce protein modifications that complicate interpretation of peptide mapping data from mass spectrometry. Considering the speed, sensitivity, and compatibility with mass spectrometry, the counterion dye stain may be more practical than any other dye-based protein stains for routine proteomic researches.
authors.affiliation: School of Pharmacy, Wenzhou Medical College, Wenzhou, Zhejiang, China.
authors.forename: Wei-Tao
authors.initials: WT
authors.lastname: Cong
authors.forename: Xu
authors.initials: X
authors.lastname: Wang
authors.forename: Sun-Young
authors.initials: SY
authors.lastname: Hwang
authors.forename: Li-Tai
authors.initials: LT
authors.lastname: Jin
authors.forename: Jung-Kap
authors.initials: JK
authors.lastname: Choi
mesh_terms: D003102:Colloids; D004396:Coloring Agents / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods / Q000592:standards; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods / Q000592:standards; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000302:isolation & purification; D005562:Formazans / Q000737:chemistry*; D057230:Limit of Detection; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D010449:Peptide Mapping / Q000379:methods; D059748:Proteolysis; D012015:Reference Standards; D012394:Rosaniline Dyes / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods / Q000592:standards; D013194:Staining and Labeling / Q000379:methods*; D014357:Trypsin / Q000737:chemistry
chemical_list: D003102:Colloids; D004396:Coloring Agents; D029968:Escherichia coli Proteins; D005562:Formazans; D010446:Peptide Fragments; D012394:Rosaniline Dyes; C009809:2-carboxy-2'-hydroxy-5'-sulfoformazylbenzene; C034887:ethyl violet; D014357:Trypsin; C004692:coomassie Brilliant Blue