SE | Serrate RNA effector molecule (rabbit antibody)

Application example



25-30 µg of total protein from Arabidopsis thaliana rosette leaves was extracted with extraction buffer containing: 100 mM Tris HCl pH 7.5, 10 % sucrose, 5 mM EDTA, 5 mM EGTA, 300 mM NaCl, 0.75 % Triton X100, 0.15 % SDS, 1 mM DTT, 1x Complete Mini EDTA-free protease inhibitor (Roche) or 7.5 % nuclear fraction obtained according to the protocol from Raczyńska et al. 2014, were separated on 10 % SDS/PAGE using semi-dry transfer and blotted 1 h to PVDF. Blots were blocked with 2.5 % milk in PBS/T for overnight at 4°C with agitation. Blot was incubated in the primary antibody at a dilution of 1: 250 in PBS-T for 1 h at RT with agitation. Blot was incubated in secondary antibody (goat anti-rabbit HRP conjugated, AS09 602, Agrisera) in 1: 5000 dlution for 1 h at RT with agitation. The blot was washed as above and developed for 5 minutes with ECL according to manufacturer's instructions. Exposure time was 600 seconds.

Courtesy of M.Sc. Mateusz Bajczyk, Department of Gene Expression, Adam Mickiewicz University, Poland