TY - JOUR
T1 - A turning point in proteome analysis
T2 - Sample prefractionation via multicompartment electrolyzers with isoelectric membranes
AU - Herbert, Ben
AU - Righetti, Pier Giorgio
PY - 2000
Y1 - 2000
N2 - Sample prefractionation, as obtained via multicompartment electrolyzers with isoelectric membranes, greatly enhanced the load ability, resolution and detection sensitivity of two-dimensional (2-D) maps in proteome analysis. This was demonstrated with different samples. In an Escherichia coli total cell extract, analysis by a 2-D map run in a pH 4-5 gradient showed many more spots when prefractionated, as compared with standard maps available in databases such as SWISS-2DPAGE. Analysis of human plasma in the pH 3-6 range showed an increase in the number of highly acidic proteins in the fractionated sample compared to whole plasma. With both samples no protein precipitation or smears occurred and much larger sample amounts could be loaded upon prefractionation, so that a large number of spots could be visualized by Coomassie staining, which is fully compatible with subsequent matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis.
AB - Sample prefractionation, as obtained via multicompartment electrolyzers with isoelectric membranes, greatly enhanced the load ability, resolution and detection sensitivity of two-dimensional (2-D) maps in proteome analysis. This was demonstrated with different samples. In an Escherichia coli total cell extract, analysis by a 2-D map run in a pH 4-5 gradient showed many more spots when prefractionated, as compared with standard maps available in databases such as SWISS-2DPAGE. Analysis of human plasma in the pH 3-6 range showed an increase in the number of highly acidic proteins in the fractionated sample compared to whole plasma. With both samples no protein precipitation or smears occurred and much larger sample amounts could be loaded upon prefractionation, so that a large number of spots could be visualized by Coomassie staining, which is fully compatible with subsequent matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis.
KW - Isoelectric membranes
KW - Prefractionation
KW - Proteome
KW - Two-dimensional maps
UR - http://www.scopus.com/inward/record.url?scp=0033672478&partnerID=8YFLogxK
U2 - 10.1002/1522-2683(200011)21:17<3639::AID-ELPS3639>3.0.CO;2-V
DO - 10.1002/1522-2683(200011)21:17<3639::AID-ELPS3639>3.0.CO;2-V
M3 - Article
C2 - 11271482
AN - SCOPUS:0033672478
SN - 0173-0835
VL - 21
SP - 3639
EP - 3648
JO - Electrophoresis
JF - Electrophoresis
IS - 17
ER -