A solubility optimization protocol for two-dimensional gel electrophoresis of cardiac tissue.
We outline a strategy for the optimization of buffer conditions for the solubilization, extraction, and isoelectric focusing (IEF) of proteins from cardiac tissue for two-dimensional gel electrophoresis (2DE). This strategy, which involves altering both the extraction and IEF buffers, allows one to ensure representation of the proteome that is as complete as possible. Initial buffer choices are given, as well as basic protocols for modifications. Although these conditions have been effectively demonstrated for human myocardium, in principle this procedure can be used for the initial screen of any new sample of tissue or cultured cells.
Stanley
Brian A
BA
Department of Physiology, Queen's University, Kingston, Ontario, Canada.
Van Eyk
Jennifer E
JE
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D007525:Isoelectric Focusing; D015394:Molecular Structure; D009206:Myocardium / Q000378:metabolism*; D020543:Proteome / Q000032:analysis; D015203:Reproducibility of Results; D012995:Solubility
D020543:Proteome