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Protein Engineering, Design and Selection
Year: 2009  |  Volume: 22  |  Issue: 10  |  Page No.: 625 - 629

Tubulin isotype specificity and identification of the epitope for antibody Tub 2.1

H Yang, F Cabral and R. Bhattacharya    


Tub 2.1, a monoclonal antibody commonly used to measure cellular tubulin content, is widely believed to recognize all β-tubulin isotypes. Though it has been used for more than two decades, the epitope for this antibody is not well established. We report for the first time that contrary to common belief, this antibody does not react with all isotypes of β-tubulin. Of the seven vertebrate β-tubulins, the more divergent class V and VI isotypes are not recognized by this antibody. Among the isotypes that do react, binding is similar for β2, β3, β4a and β4b but lower for β1, the most abundant isotype. Expression of chimeric tubulins verified that the epitope for Tub 2.1 is near the C-terminal end of β-tubulin. Site-directed mutagenesis of this region in nonreactive β5 to match the sequence of β4b resulted in strong reaction to Tub 2.1 and narrowed the epitope to amino acids 431–436.

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