Postfractionation for enhanced proteomic analyses: routine electrophoretic methods increase the resolution of standard 2D-PAGE.
Here we have addressed common issues of resolution in two-dimensional polyacrylamide gel electrophoresis (2DE) experiments including proteins 'stacked' at pH extremes, unresolved peptides migrating at the front of separation, and areas of the 2D gel obscured by high abundance proteins. Postfractionation, by selective application of well-established electrophoretic separations immediately following standard 2DE, yields markedly improved resolution in these traditional problem areas using no more specialized equipment or techniques than SDS-PAGE itself.
Butt
R Hussain
RH
Department of Physiology, Hotchkiss Brain Institute, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada.
Coorssen
Jens R
JR
D000818:Animals; D001923:Brain Chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000592:standards*; D004591:Electrophoresis, Polyacrylamide Gel; D008099:Liver / Q000737:chemistry; D051379:Mice; D008815:Mice, Inbred Strains; D009206:Myocardium / Q000737:chemistry; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*
D011506:Proteins