Anti-RabG | Ras-related protein RABG (global antibody)

Protein extracted freshly, from Wild Type (Col-0) Arabidopsis thaliana and mixed with 2x Laemmli sample buffer plus beta-mercaptoethanol and denatured at 95 °C for 5 min prior to SDS-PAGE analysis. Samples were separated on 4-20% SDS-PAGE at room temperature and blotted for 7 min to nitrocellulose Membrane (pore size of 0.2 µm), using: semi-wet transfer system. Blot was blocked with 1x TBS 1% Casein Blocker for 1h/RT. Blot was incubated in the primary antibody at a dilution of 1: 500 for ON/4°C with agitation. The antibody solution was decanted, then washed 3 times for 5 min in TBS-T at RT. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1: 25k for 3h/RT with agitation. The blot was washed as above and developed with a following chemiluminescent detection reagent: SuperSignal West Pico Plus chemiluminescent ThermoScientific). Exposure time was a few seconds to 1 minutes.

Courtesy of Sahar Farami, University of Glasgow, UK

Alternative names: Ras-related protein Rab71 (AtRab71), Ras-related protein Rab7B (AtRab7B)