Purification of brain tubulin through two cycles of polymerization-depolymerization in a high-molarity buffer

Protein Expr Purif. 2003 Nov;32(1):83-8. doi: 10.1016/S1046-5928(03)00218-3.


Microtubules can be assembled in vitro from purified alpha/beta tubulin heterodimers in the presence of GTP. Tubulin is routinely obtained from animal brain tissue through repetitive cycles of polymerization-depolymerization, followed by ion-exchange chromatography to remove any contaminating microtubule-associated proteins and motors. Here, we show that only two cycles of polymerization-depolymerization of pig brain tubulin in the presence of a high-molarity PIPES buffer allow the efficient removal of contaminating proteins and production of a high-concentration tubulin solution. The proposed protocol is rapid and yields more active tubulin than the traditional ion-exchange chromatography-based procedures.

MeSH terms

  • Animals
  • Biopolymers / chemistry
  • Biopolymers / isolation & purification*
  • Biopolymers / metabolism*
  • Brain Chemistry*
  • Buffers
  • Cellulose / analogs & derivatives*
  • Chromatography, Liquid
  • Microtubules / chemistry
  • Microtubules / metabolism
  • Protein Structure, Quaternary
  • Swine
  • Tubulin / chemistry
  • Tubulin / isolation & purification*
  • Tubulin / metabolism*


  • Biopolymers
  • Buffers
  • Tubulin
  • Cellulose
  • phosphocellulose