Fig. 6. The specificity of anti-twitchin D2 antibody. (A) Immunoblotting using the anti-twitchin D2 antibody against ABRM myofibrillar proteins. Lane 1, protein-loaded gel stained with Commassie Brilliant Blue; lane 2, antibody loaded onto the PVDF membrane reacted only with twitchin and not to the major components of thick filaments, myosin heavy chain, myorod and paramyosin. (B) Immunoblotting using the anti-twitchin D2 antibody against TWD2-S and its phosphorylated form (P-TWD2-S). Arrowheads indicate twitchin peptide and the positions of molecular mass markers (kDa) are shown. (C) Differences in the reactivity of the anti-twitchin D2 antibody against TWD2-S and P-TWD2-S. Closed and open circles represent TWD2-S and P-TWD2-S, respectively.