Antibody Dilution and Detection Protocol

Antibodies used in this study were diluted in 5% w/v bovine serum albumin in TBS supplemented with 0.1% Tween-20 (TBS-T) and 0.03% w/v sodium azide. The rabbit monoclonal antibody for total LRRK2 (N-terminus) was purified at the University of Dundee (16 (link)). Anti-GFP (PABG1; Chromotek, used at 1:1000), anti-HA (3F10; Merck, used at 1:1000), anti-pT72-Rab8a (MJF-R20; Abcam, used at 0.5 μg/mL), anti-LRRK2 C-terminal (N241A/34; Neuromab, used at 1:1000), and anti-α-tubulin (3873S; CST, used at 1:5000). Secondary antibodies used were LI-COR IRDye for 800CW goat anti-rabbit (925–32211), goat anti-mouse (926–32210), and 680LT goat anti-rat (925–68029) and goat anti-mouse (926–68020), all used at 1:10,000 dilution in TBS-T.