Electrophoretic separation and quantitation of cardiac myosin heavy chain isoforms in eight mammalian species

The American Journal of Physiology
Peter J Reiser, W O Kline

Abstract

A protocol for sample preparation and gel electrophoresis is described that reliably results in the separation of the alpha- and beta-isoforms of cardiac myosin heavy chain (MHC-alpha and MHC-beta) in eight mammalian species. The protocol is based on a simple, nongradient denaturing gel. The magnitude of separation of MHC-alpha and MHC-beta achieved with this protocol is sufficient for quantitative determination of the relative amounts of these two isoforms in mouse, rat, guinea pig, rabbit, canine, pig, baboon, and human myocardial samples. The sensitivity of the protocol is sufficient for the detection of MHC isoforms in samples at least as small as 1 microgram. The glycerol concentration in the separating gel is an important factor for successfully separating MHC-alpha and MHC-beta in myocardial samples from different species. The effect of sample load on MHC-alpha and MHC-beta band resolution is illustrated. The results also indicate that inclusion of a homogenization step during sample preparation increases the amount of MHC detected on the gel for cardiac samples to a much greater extent than for skeletal muscle samples. Although the protocol described in this study is excellent for analyzing cardiac samples, it should be...Continue Reading

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Citations

Oct 18, 2011·Journal of Comparative Physiology. B, Biochemical, Systemic, and Environmental Physiology·Michelle R SheroJennifer M Burns
Jun 1, 2014·American Journal of Physiology. Heart and Circulatory Physiology·Peter J Reiser, Christine S Moravec
Apr 19, 2002·Journal of Applied Physiology·Hao-Jie WangTai Yao
Mar 4, 2000·Circulation Research·S MiyataL A Leinwand
Jan 6, 2004·American Journal of Physiology. Heart and Circulatory Physiology·Motoyuki IemitsuIwao Yamaguchi
Sep 18, 2002·American Journal of Physiology. Heart and Circulatory Physiology·Norman R AlpertDavid M Warshaw
Mar 21, 2007·American Journal of Physiology. Heart and Circulatory Physiology·Aziz GuellichCatherine Coirault
Jul 4, 2020·American Journal of Physiology. Heart and Circulatory Physiology·Paul RobinsonCharles Redwood
Mar 15, 2001·American Journal of Physiology. Heart and Circulatory Physiology·P J ReiserC Schomisch Moravec
Aug 23, 2003·American Journal of Physiology. Heart and Circulatory Physiology·Yan ZhongMohammed A Matlib
May 11, 2004·Journal of Applied Physiology·Takeshi HashimotoSadayoshi Taguchi
Dec 24, 2005·American Journal of Physiology. Cell Physiology·Mark BergrinPeter J Reiser
Oct 9, 2004·American Journal of Physiology. Heart and Circulatory Physiology·Veronica L M RundellPieter P de Tombe
Feb 19, 2008·American Journal of Physiology. Heart and Circulatory Physiology·Bradley M PalmerDavid W Maughan
May 18, 2001·Journal of Applied Physiology·T L ClantonN R Prabhakar
Mar 8, 2005·Journal of Muscle Research and Cell Motility·Sabahattin Bicer, Peter J Reiser
Jun 29, 2004·Journal of Applied Physiology·Cynthia A CarnesPeter J Reiser
Jun 19, 2012·American Journal of Physiology. Heart and Circulatory Physiology·Mellani LeftaKaryn A Esser
Oct 23, 2004·American Journal of Physiology. Heart and Circulatory Physiology·Bradford C BluntPolly A Hofmann
Sep 10, 2003·The Journal of Physiology·Norio FukudaHenk Granzier
Jul 1, 2020·Applied Physiology, Nutrition, and Metabolism = Physiologie Appliquée, Nutrition Et Métabolisme·Kevin BoldtWalter Herzog
Jan 4, 2021·Journal of Muscle Research and Cell Motility·Galina V KopylovaDaniil V Shchepkin
Sep 22, 2020·Mechanisms of Ageing and Development·Kevin BoldtWalter Herzog
Jan 19, 2021·Biochemical and Biophysical Research Communications·Anastasia KhokhlovaDaniil Shchepkin
Feb 9, 2021·Computers in Biology and Medicine·Robson Rodrigues da SilvaRosana Almada Bassani
Apr 8, 2021·Journal of Muscle Research and Cell Motility·Ramzi A Al-HoraniMohammed Ihsan

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