Isoelectric focusing in long immobilized pH gradient gels to improve protein separation in proteomic analysis

Julia Poland, Michael Cahill, Pranav Sinha

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The number of protein spots detected on two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) gels increases as the gel size increases. The largest commercially available systems resolve a few thousand spots, being only a fraction of the total proteome. We have developed an extremely long isoelectric focusing (IEF) system aimed at more complete protein profiling. The system is especially well suited to sensitive detection methods, such as radioactive detection. The major constraint preventing progress in this area has been the inability to create an even density gradient during the immobilized pH gradient (IPG) casting process. We demonstrate for the first time that this constraint can be effectively overcome, to enable greatly increased IEF separating power with all the advantages of IPG technology
Original languageEnglish
Pages (from-to)1271-1275
Number of pages5
JournalElectrophoresis
Volume24
Issue number7-8
DOIs
Publication statusPublished - 2003

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