Quantification of acetylated tubulin

CHESTA, M.E.; CARBAJAL, A.; BISIG, C. G.; ARCE, C.A.

WILEY-LISS, DIV JOHN WILEY & SONS INC

Lugar: New York; Año: 2013 vol. 70 p. 297 - 297

he acetylation/deacetylation of Lys40 of the α-subunit is an important post-translational modification undergone by tubulin during the life of a cell. Many previous studies have addressed the physiological role of this acetylation process using various approaches based on changes of acetylated tubulin (AcTubulin) content. In most of these studies, however, the actual amounts of AcTubulin were not known and it was difficult to draw conclusions. We present here a simple method to estimate the percentage of AcTubulin relative to total tubulin. The method is based on acetylation of the tubulin sample with acetic anhydride, Western blotting stained by anti-AcTubulin antibody, and comparison of the optical density of the AcTubulin band with that of a corresponding sample that was not chemically acetylated.