Journal: Molecular Neurodegeneration
Article Title: Tau phosphorylation by GSK-3? promotes tangle-like filament morphology
Figure Lengend Snippet: Tau phosphorylation by GSK-3B . A) SDS-PAGE analysis of tau protein incubated for 20 h in the absence (lanes 1, 4, 7, 10, and 13) or the presence of 0.006 U GSK-3β per pmol tau (lanes 2, 5, 8, 11, 14) or 0.018 U GSK-3β per pmol tau (lanes 3, 6, 9, 12, and 15). A definite band shift in the migration of phosphorylated tau can be detected with increasing time and kinase concentration. B) The amount of γ- 32 P incorporation over time using 0.018 U GSK-3β per pmol tau (open circles, right y-axis) is compared to the SDS-PAGE analysis in Panel A (open squares, left y-axis). Lines are drawn through the points to ease comparison. Data represents a single trial.
Article Snippet: Determination of phosphate incorporation Tau protein at a final concentration of 16 μM was incubated with 0.018 U GSK-3β per pmol tau in buffer containing 40 mM HEPES, pH 7.64, 5 mM EGTA, 3 mM MgCl2 , and 2 mM ATP containing 10 μCi [γ-32 P] labeled ATP (Specific activity: 3000 Ci/mmol) (Perkin-Elmer, Boston, MA) for 20 h at 30°C.
Techniques: SDS Page, Incubation, Electrophoretic Mobility Shift Assay, Migration, Concentration Assay