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

被引:27
作者
Poland, J
Cahill, MA
Sinha, P
机构
[1] ProteoSys Ag, D-55129 Mainz, Germany
[2] Humboldt Univ, Fac Med, Klinikum Charite, Inst Lab Med & Pathobiochem, Berlin, Germany
关键词
immobilized pH gradient; isoelectric focusing; two-dimensional polyacrylamide gel electrophoresis;
D O I
10.1002/elps.200390163
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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,
引用
收藏
页码:1271 / 1275
页数:5
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