Analysis of differential detergent fractions of an AtT-20 cellular homogenate using one- and two-dimensional capillary electrophoresis.
Differential detergent fractionation was used to sequentially extract cytosolic, membrane, nuclear, and cytoskeletal fractions from AtT-20 cells. Extracted components were denatured by sodium dodecyl sulfate (SDS) and then labeled with the fluorogenic reagent 3-(2-furoyl) quinoline-1-carboxaldehyde. Both capillary sieving electrophoresis (CSE) and micellar electrokinetic capillary chromatography (MECC) were used to separate labeled components by one-dimensional (1D) electrophoresis. Labeled components were also separated by two-dimensional (2D) capillary electrophoresis; CSE was employed in the first dimension and MECC in the second dimension. Roughly 150 fractions were transferred from the first to the second capillary for this comprehensive analysis in 2.5 h.
Fazal
Md Abul
MA
Department of Chemistry, University of Washington, Seattle, 98195-1700, USA.
Palmer
Vanessa R
VR
Dovichi
Norman J
NJ
D000818:Animals; D002478:Cells, Cultured; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods; D003902:Detergents / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D051379:Mice; D011506:Proteins / Q000032:analysis
D003902:Detergents; D011506:Proteins