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Cardiac muscle protein analysis by high‐resolution and microscale two‐dimensional gel electrophoresis
Author(s) -
Tanaka Takao,
Kawamura Keishiro
Publication year - 1988
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150090207
Subject(s) - cardiac muscle , myosin , tropomyosin , electrophoresis , isoelectric focusing , agarose , chromatography , chemistry , actin , myoglobin , troponin , two dimensional gel electrophoresis , resolution (logic) , biochemistry , biology , anatomy , proteomics , psychology , artificial intelligence , psychiatry , myocardial infarction , gene , computer science , enzyme
An improved method of two‐dimensional gel electrophoresis of small muscle samples is described. Isoelectric focusing of cardiac whole muscle homogenate in agarose gels containing urea and detergent has a markedly increased resolution. Equilibration of the first‐dimensional gels with detergent before application to the second‐dimensional gels is unnecessary in this system. By applying this method to rat cardiac whole muscle, high‐molecular weight proteins, such as myosin heavy chains, are focused on the first‐dimensional gels and, in addition, minor components are resolved on the second‐dimensional gels, without loss during equilibration with detergent. The two‐dimensional electrophoretic patterns of rat cardiac whole muscle obtained with this method reveal numerous clearly separated spots. By analyzing the two‐dimensional electrophoretic patterns of rat cardiac whole muscle and various rat cardiac fractions, and by staining the calcium‐binding proteins with “Stains‐all”, we identified some cardiac muscle components, such as myosin heavy and light chains, actin, tropomyosin, and troponin C, but additional work is required to identify the remaining spots. The two‐dimensional electrophoretic system described here makes possible the effective resolution of whole cardiac muscle homogenate from small samples, and looks promising as an aid to muscle research.