Protocol for high‐resolution electrophoresis separation of myosin heavy chain isoforms in bovine skeletal muscle

Abstract

In this short communication we describe a specific protocol for SDS-PAGE separation of adult bovine myosin heavy-chain (MyHC) isoforms. The conditions defined in this protocol allow a good separation with a good reproducibility of the four MyHC isoforms (MyHC I, IIa, IIx, IIb) identified in adult skeletal muscle of this species. This procedure uses mini-gel electrophoresis system and does not involve preparation of gradient separating gels. In addition, this protocol can also be applied to the electrophoretic separation of ovine and camel MyHC isoforms.

Cardiomyopathy Mutations in the Tail of β-Cardiac Myosin Modify the Coiled-coil Structure and Affect Integration into Thick Filaments in Muscle Sarcomeres in Adult Cardiomyocytes

“…Levels of eGFP-MHC expression were compared with endogenous levels of MHC by SDS gel analysis using a protocol that separates MHC isoforms in a minigel system (28). Briefly, the separating gel contained 35% v/v glycerol, 9% total acrylamide of which 1/49 was bisacrylamide, 230 mM Tris-HCl, 115 mM glycine, pH 8.8, and 0.4% w/v SDS; and the stacking gel contained 47% v/v glycerol, 6% w/v acrylamide (1/49 bisacrylamide), 110 mM Tris-HCl, 6 mM EDTA, pH 6.8, and 0.4% w/v SDS.…”

Sensory quality of meat from eight different types of cattle in relation with their biochemical characteristics

“…Following centrifugation, the supernatant was diluted 1:1 (v/w) with glycerol at 87% and PAGE electrophoresis according to (Picard et al 2011) using 9.2% polyacrylamide gels which 402 were made in the Bio-Rad mini-Protean II Dual slab cell system. After staining, the gels were LT muscle taken 24 h after slaughter as previously described (Karamichou et al 2007;415 Gagaoua et al 2013).…”

Understanding Early Post-Mortem Biochemical Processes Underlying Meat Color and pH Decline in the Longissimus thoracis Muscle of Young Blond d’Aquitaine Bulls Using Protein Biomarkers

“…Electrophoresis and quantification of Myosin Heavy Chain (MyHC) isoforms145 Three other biomarkers corresponded to MyHC and were determined according to Picard 146 et al24 using an adequate SDS-PAGE and expressed in percentage. Briefly, 100 mg of frozen 147 muscle were ground in 5mL of extraction buffer solution containing 0.5 M NaCl, 20 mM Na 148 Pyrophosphate, 50 mM Tris, 1 mM EDTA and 1 mM Dithiothreitol.…”