Two-dimensional gel electrophoresis for the identification of signaling targets.
Two-dimensional electrophoresis (2-DE) is a powerful technique to differentially display patterns of protein expression and posttranslational modifications, providing a good strategy to monitor molecular responses induced by the activation or inactivation of specific signaling pathways. In this chapter, optimized protocols for 2-DE using extracts from tissue culture are provided. Protocols for in-gel digestion of gel-resolved proteins, which allow protein identification by mass spectrometry are also discussed.
Kabuyama
Yukihito
Y
Department of Chemistry and Biochemistry, Howard Hughes Medical Institute, University of Colorado at Boulder, USA.
Polvinen
Kirsi K
KK
Resing
Katheryn A
KA
Ahn
Natalie G
NG
D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D007525:Isoelectric Focusing; D013058:Mass Spectrometry; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction; D016622:Silver Staining / Q000379:methods
D011506:Proteins