Purification of bovine Tau versions by affinity chromatography.
The nervous-system-specific microtubule-associated Tau proteins promote microtubule stability and assembly. Tau is transiently phosphorylated at about 30 positions and additionally O- and N-glycosylated. Bovine Tau was prepared from a calf brain and purified by affinity chromatography using immobilized monoclonal antibody (mAb) BT-2, which recognizes all Tau-splicing isoforms. Tau was obtained in high purities well above 90% containing even highly phosphorylated Tau versions with isoelectric points below pH 5 without discrimination. Moreover, these highly phosphorylated versions were detected only after purification. The purification progress and the final purity were studied by two-dimensional gel electrophoresis (2DE) using both protein and phosphoprotein stains as well as immunoblots.
Volke
Daniela
D
Bioanalytics, Center for Biotechnology and Biomedicine (BBZ), Faculty of Chemistry and Mineralogy, University of Leipzig, Deutscher Platz 5, 04103 Leipzig, Germany.
Hoffmann
Ralf
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000937:Antigen-Antibody Reactions; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006863:Hydrogen-Ion Concentration; D012680:Sensitivity and Specificity; D016875:tau Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D016875:tau Proteins