title: Quantification of acetylated tubulin.
abstract: The acetylation/deacetylation of Lys40 of the α-subunit is an important posttranslational 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 antiAcTubulin antibody, and comparison of the optical density of the AcTubulin band with that of a corresponding sample that was not chemically acetylated.
authors.affiliation: Departamento de Química Biológica, Facultad de Ciencias Químicas, Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC), UNC-CONICET, Universidad Nacional de Córdoba, Córdoba, Argentina.
authors.forename: María E
authors.initials: ME
authors.lastname: Chesta
authors.forename: Agustín
authors.initials: A
authors.lastname: Carbajal
authors.forename: C Gastón
authors.initials: CG
authors.lastname: Bisig
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Arce
mesh_terms: D000107:Acetylation; D000818:Animals; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002623:Chemistry Techniques, Analytical; D004591:Electrophoresis, Polyacrylamide Gel; D051379:Mice; D008870:Microtubules / Q000737:chemistry*; D014404:Tubulin / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D014404:Tubulin
keywords: acetylated tubulin; chemical acetylation; microtubules; quantification method