Thanks you very much Mohamed. I need a specific antibody for Western Blot. I want to study the separate isoforms (2a, 2x, I), however I have a little problem because I didn't find a comercial specific antibody for myosin heavy chain 2x. Do you know a company that sell a specific antibody for 2x in bovine muscle using Western Blot?

My lab has developed Mabs against 2B and 2B MyHCs (Lucas CA, Kang LH, and Hoh JF. Monospecific antibodies against the three mammalian fast limb myosin heavy chains. Biochem Biophys Res Commun 272: 303-308, 2000). They work even in marsupials (Zhong WW, Lucas CA, Kang LH, and Hoh JF. Electrophoretic and immunochemical evidence showing that marsupial limb muscles express the same fast and slow myosin heavy chains as eutherians. Electrophoresis 22: 1016-1020, 2001), and should work in bovine tissues. They are MAbs 6H1 and 10F5 (available from Developmental Studies Hybridoma Bank; University of Iowa, IA), specific to 2X and 2B MyHCs, respectively. They work in Westerns. Good luck with your work, Cruz.

My lab has developed Mabs against 2B and 2B MyHCs (Lucas CA, Kang LH, and Hoh JF. Monospecific antibodies against the three mammalian fast limb myosin heavy chains. Biochem Biophys Res Commun 272: 303-308, 2000). They work even in marsupials (Zhong WW, Lucas CA, Kang LH, and Hoh JF. Electrophoretic and immunochemical evidence showing that marsupial limb muscles express the same fast and slow myosin heavy chains as eutherians. Electrophoresis 22: 1016-1020, 2001), and should work in bovine tissues. They are MAbs 6H1 and 10F5 (available from Developmental Studies Hybridoma Bank; University of Iowa, IA), specific to 2X and 2B MyHCs, respectively. They work in Westerns. Good luck with your work, Cruz.

Thanks you very much JosepH.

The Schiaffino's antibodies against type 1, 2A and 2B myosins, which can be found at Developmental Studies Hybridoma Bank like the ones proposed by Joseph, should work as well.

They are called BA-D5, SC71 and BF-F3 respectively.

In the attached link you can find a discussion in which I posted a couple of protocols.

https://www.researchgate.net/post/Does_anybody_have_experience_with_muscle_fiber_typing_Any_suggestions#view=5670a21164e9b244858b4578

Hello Dr. Stefano Cicilio. Thank you very much for your answer. I'm working com western blot in bovine muscle and really I'm having problems with 2x isoform. I tested 6H1 antibody of Hybridoma bank but 2x isoform wasn't identified. I don't know if my problem was in the extraction process. I used modified radioimmunoprecipitation buffer (RIPPA) and TRITON. How you do the extraction process?. Thanks, very much.