Fig. 8. Purification of microtubules from Sf9 cells expressing the truncated forms of MAP2 and full-length MAP2b and MAP2c. Microtubules were prepared 64 hours following the infection as described in Materials and Methods. (A) The purity of microtubules was verified by Coomassie staining; 2 µg of microtubules were loaded per lane. Coomassie staining revealed microtubules as well as their associated protein. The numbers indicate MAP2b, MAP2c, MAP2b-1, MAP2b-2, MAP2b-3 and Mt, respectively. The molecular weight standards are (top to bottom): myosin (203 kDa), ß-galactosidase (135 kDa), bovin serum albumin (86 kDa) and carbonic anhydrase (42 kDa). On a 7.5% acrylamide gel, Mt and tubulin migrate at the same level and thus it is not possible to visualize Mt. However, Mt was revealed by dot blotting using the 46.1 antibody. (B) To verify the level of tubulin, proteins were transferred on a nitrocellulose membrane. The membrane was revealed with an anti-tubulin monoclonal antibody.

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