Isolation of organelles and prefractionation of protein extracts using free-flow electrophoresis.
One of the major obstacles in the analysis of proteomes is the extreme complexity of any particular cell or biological fluid. Free-flow electrophoresis (FFE) is a powerful tool for reduction of this complexity, which is a prerequisite for systematic and comprehensive protein analyses. Protocols are provided in this unit for sample fractionation at two different stages: on the protein level by isoelectric focusing FFE fractionation of crude protein mixtures such as whole cell lysates, and on a subcellular level by zone-electrophoretic FFE purification of organelles.
Weber
Peter J A
PJA
Tecan Munich GmbH, Kirchheim, Germany.
Weber
Gerhard
G
Tecan Munich GmbH, Kirchheim, Germany.
Eckerskorn
Christoph
C
Tecan Munich GmbH, Kirchheim, Germany.
D004586:Electrophoresis / Q000295:instrumentation / Q000379:methods; D007525:Isoelectric Focusing / Q000379:methods; D015388:Organelles / Q000737:chemistry*; D011506:Proteins / Q000032:analysis*
D011506:Proteins