Rapid subpicogram protein detection on a microchip without denaturing.
Protein size separation based on sodium dodecyl sulfate-gel electrophoresis (SDS-GE) requires denaturing, but we propose that denaturing is unnecessary for analysis by microchip electrophoresis (micro-CE). By omitting the protein denaturing process, we achieved not only shortened total analysis time, but also dramatically improved sensitivity without compromising size determination. The detection limit was improved to 0.1 ng/microL under conditions without denaturing and 600 pg (9.0 femtomol) of bovine serum albumin was detectable, which equals levels detectable by Silver stain, although a routine method by microchips in the Coomassie Blue detection level.
Tabuchi
Mari
M
Department of Medicinal Chemistry, Faculty of Pharmaceutical Sciences, The University of Tokushima, Tokushima, Japan. tabuchi@ph.tokushima-u.ac.jp
Kuramitsu
Yasuhiro
Y
Nakamura
Kazuyuki
K
Baba
Yoshinobu
Y
D030562:Databases, Protein; D004586:Electrophoresis / Q000379:methods; D006360:Heat-Shock Proteins / Q000032:analysis; D006801:Humans; D019169:Jurkat Cells / Q000737:chemistry; D013058:Mass Spectrometry; D008832:Microchemistry / Q000379:methods*; D011489:Protein Denaturation; D011506:Proteins / Q000032:analysis*; D020543:Proteome / Q000032:analysis; D012680:Sensitivity and Specificity
D006360:Heat-Shock Proteins; D011506:Proteins; D020543:Proteome