Detection of endogenous S1292 LRRK2 autophosphorylation in mouse tissue as a readout for kinase activity

Characterization of pS1292 LRRK2 antibody. a Increasing amounts of HEK293FT lysates transiently expressing WT LRRK2 (A) or S1292A LRRK2 (B) were analyzed by western blot and probed with the commercially available anti-pS1292 LRRK2 antibody (ab203181). The antibody showed minimal non-specific bands and linear detection in the range tested (b) (n？=？3 blots; mean with SD bars shown). Transiently expressed LRRK2 genetic variants were probed for S1292 phosphorylation (c, d). Strong pS1292 signal was detected in the G2019S and I2020T kinase hyperactive variants and no signal detected at the S1292A phospho-null and K1906M kinase-dead variant. The R1441C and Y1699C variants showed increased S1292 phosphorylation compared to WT which is in accordance to previous literature (c, d) (n？=？4; one-way ANOVA with Tukey’s post hoc test; F(8, 27)？=？210.1