|Wu, Xia - University Of Washington|
|Oh, Man-ho - Chungnam National University|
Submitted to: Bio-protocol
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/1/2013
Publication Date: 10/5/2013
Citation: Wu, X., Clough, S.J., Huber, S.C., Oh, M. 2013. EF-2DE Analysis and Protein Identification. Bio-protocol. 3(19):1-8.
Interpretive Summary: We described the technique isoelectric focusing followed by SDS-PAGE (IEF-2DE), which separates proteins for clearer identification. The technique is particularly useful to distinguish different forms of the same protein, as well as various modifications of a protein. This technique will be useful for molecular biologists.
Technical Abstract: Isoelectric focusing followed by SDS-PAGE (IEF-2DE) separates proteins in a two-dimensional matrix of protein pI (Protein Isoelectric Point) and molecular weight (MW). The technique is particularly useful to distinguish protein isoforms (Radwan et al., 2012) and proteins that contain post-translational modifications such as phosphorylation (Oh et al., 2012) and lysine acetylation (Wu et al., 2011). Proteins that are separated by IEF-2DE can be identified by immunoblot analysis using sequence-specific antibodies or by mass spectrometry.