A simple procedure for obtaining large quantities of tubulin subunits from newborn mouse brain.
A simple, efficient two-step procedure of DEAE--Sephadex A--50 column chromatography followed by hydroxyapatite column chromatography for purification of alpha- and beta-tubulin subunits from newborn mouse brain is described. Stored frozen mouse brains can be used as a convenient starting material. Differential elution of alpha and beta subunits from hydroxyapatite was achieved using a linear gradient of 0.2--0.3 M phosphate buffer containing 2 M urea, 0.1% sodium dodecyl sulphate and 1 mM dithiothreitol. 20 mg of alpha-tubulin (purity: greater than 85%) and 16 mg of beta-tubulin (purity greater than 95%) completely separated from each other can be obtained in one experiment.
D000818:Animals; D000831:Animals, Newborn; D001923:Brain Chemistry; D002845:Chromatography / Q000379:methods; D005260:Female; D006023:Glycoproteins / Q000302:isolation & purification*; D008297:Male; D051379:Mice; D014404:Tubulin / Q000302:isolation & purification*
D006023:Glycoproteins; D014404:Tubulin