Premium
Isoelectric focusing in long immobilized pH gradient gels to improve protein separation in proteomic analysis
Author(s) -
Poland Julia,
Cahill Michael A.,
Sinha Pranav
Publication year - 2003
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200390163
Subject(s) - isoelectric focusing , immobilized ph gradient , chromatography , proteome , polyacrylamide , chemistry , polyacrylamide gel electrophoresis , protein purification , two dimensional gel electrophoresis , separation process , fraction (chemistry) , proteomics , biochemistry , polymer chemistry , gene , enzyme
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,