title: Enhanced protein electrophoresis technique for separating human skeletal muscle myosin heavy chain isoforms.
abstract: Talmadge and Roy (J. Appl. Physiol. 1993, 75, 2337-2340) previously established a sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) protocol for separating all four rat skeletal muscle myosin heavy chain (MHC) isoforms (MHC I, IIa, IIx, IIb); however, when applied to human muscle, the type II MHC isoforms (Ila, IIx) are not clearly distinguished. In this brief paper we describe a modification of the SDS-PAGE protocol which yields distinct and consistent separation of all three adult human MHC isoforms (MHC I, IIa, IIx) in a minigel system. MHC specificity of each band was confirmed by Western blot using three monoclonal IgG antibodies (mAbs) immunoreactive against MHCI (mAb MHCs, Novacastra Laboratories), MHCI+IIa (mAb BF-35), and MHCIIa+IIx (mAb SC-71). Results provide a valuable SDS-PAGE minigel technique for separating MHC isoforms in human muscle without the difficult task of casting gradient gels.
authors.affiliation: Department of Human Studies, University of Alabama at Birmingham 35294-1250, USA. mbamman@uab.edu
authors.forename: M M
authors.initials: MM
authors.lastname: Bamman
authors.forename: M S
authors.initials: MS
authors.lastname: Clarke
authors.forename: R J
authors.initials: RJ
authors.lastname: Talmadge
authors.forename: D L
authors.initials: DL
authors.lastname: Feeback
mesh_terms: D000328:Adult; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D018482:Muscle, Skeletal / Q000737:chemistry; D018995:Myosin Heavy Chains / Q000302:isolation & purification*; D020033:Protein Isoforms / Q000302:isolation & purification
chemical_list: D020033:Protein Isoforms; D018995:Myosin Heavy Chains