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Enhanced protein electrophoresis technique for separating human skeletal muscle myosin heavy chain isoformsTalmadge 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.
Document ID
20040141985
Document Type
Reprint (Version printed in journal)
Authors
Bamman, M. M. (University of Alabama at Birmingham 35294-1250 United States)
Clarke, M. S.
Talmadge, R. J.
Feeback, D. L.
Date Acquired
August 22, 2013
Publication Date
March 1, 1999
Publication Information
Publication: Electrophoresis
Volume: 20
Issue: 3
ISSN: 0173-0835
Subject Category
Life Sciences (General)
Funding Number(s)
CONTRACT_GRANT: M01 RR00073
Distribution Limits
Public
Copyright
Other
Keywords
NASA Center JSC
NASA Discipline Musculoskeletal