Solution phase isoelectric fractionation in the multi-compartment electrolyser: a divide and conquer strategy for the analysis of complex proteomes.
Sample complexity frequently interferes with the analysis of low-abundance proteins by two-dimensional gel electrophoresis (2DGE). Ideally, high abundance proteins should be removed, allowing low-abundance proteins to be applied at much higher concentrations than is possible with the unfractionated sample. One approach is to partition the sample in a manner that segregates the bulk of extraneous proteins from the protein(s) of interest. Solution phase isoelectric focusing in the multi-compartment electrolyser generates fractions of discrete isoelectric point (pI) intervals allowing isolated narrow segments of a proteome to be analysed individually by 2DGE. It is particularly useful for the isolation of low-abundance proteins of extremely basic or acidic pI.
Smejkal
Gary B
GB
Proteome Systems, 14 Gill Street, Woburn, MA 01801, USA. smejkal@proteomesystems.com
Lazarev
Alexander
A
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007091:Image Processing, Computer-Assisted; D007525:Isoelectric Focusing / Q000295:instrumentation* / Q000379:methods*; D008566:Membranes; D020543:Proteome; D040901:Proteomics / Q000295:instrumentation* / Q000379:methods*
D029968:Escherichia coli Proteins; D020543:Proteome