title: Protocol for high-resolution separation of rodent myosin heavy chain isoforms in a mini-gel electrophoresis system.
abstract: Skeletal muscle comprises several fiber types classified based on their contractile and metabolic properties. Skeletal muscle fiber types are classified according to their myosin heavy chain isoforms (MyHC I, IIa, IIx, and IIb). We attained good separation of MyHC isoforms in a mini-gel system by modifying a previously developed electrophoresis protocol. Increased glycerol and decreased cross-linking agent concentrations improved the separation of MyHC isoforms. Sample preparation with dithiothreitol and protease inhibitors produced clear MyHC band boundaries. This protocol included silver staining, with a linear range. The protocol provided high resolution and a highly accurate assay of rodent MyHC isoforms.
authors: Department of Bioscience and Biotechnology, Graduate School of Agriculture, Kyushu University, Higashi-ku, Fukuoka 812-8581, Japan. wataru_m@agr.kyushu-u.ac.jp
authors: Wataru
authors: W
authors: Mizunoya
authors: Jun-ichiro
authors: J
authors: Wakamatsu
authors: Ryuichi
authors: R
authors: Tatsumi
authors: Yoshihide
authors: Y
authors: Ikeuchi
mesh_terms: D020106:Acrylamide / Q000737:chemistry; D000818:Animals; D003432:Cross-Linking Reagents / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005990:Glycerol / Q000737:chemistry; D051379:Mice; D018995:Myosin Heavy Chains / Q000032:analysis* / Q000737:chemistry*; D020033:Protein Isoforms / Q000032:analysis* / Q000737:chemistry; D051381:Rats; D015203:Reproducibility of Results; D016622:Silver Staining
chemical_list: D003432:Cross-Linking Reagents; D020033:Protein Isoforms; D020106:Acrylamide; D018995:Myosin Heavy Chains; D005990:Glycerol