Nondenaturing electrophoresis as a tool to investigate tubulin complexes

A protein molecule may exist as a monomer, homo-oligomer, or hetero-oligomer in a multiprotein complex. One-dimensional (1-D) native electrophoresis has long been used to characterize tubulins and their complexes. In this chapter, we describe the simplest way to identify the state of aggregation of commercial or homemade tubulins for further studies based on 1-D electrophoresis under nondenaturing conditions. We present a series of detailed protocols that can be used to analyze the maturation of alpha- and beta-tubulins and to identify the complexes formed during the folding and dimerization pathway as well as their stability.