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Figure 3


Fig. 3. (A) Coomassie-Blue-stained agarose gel of rat muscle (Rm), locust muscle (Lm) and crane-fly muscle (CFm). The highest molecular mass components in crane-fly and locust muscle (arrows) run about half-way between rat titin and rat nebulin, similarly to the doublets of mini-titin (Nave and Weber, 1990). The arrowhead indicates the band used for mass spectrometry analysis. (B) Western blots of locust muscle probed with three anti-D-titin antibodies. All three antibodies stain the highest molecular weight bands (arrows) seen in the Coomassie-Blue-stained gels, corresponding to mini-titin, as indicated by their position relative to rat titin and rat nebulin, which act as markers (Warren et al., 2003). Myosin runs with the Bromophenol Blue dye front and is at the bottom of the lanes. The proteins were separated on agarose gels, transferred to PVDF membrane and visualized with ECL. The same membrane was used for all three anti-D-titin antibodies (after stripping and reprobing).





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