Anti-FER | Receptor-like protein kinase FERONIA

50 µg/well of total protein extracted freshly from Arabidopsis thaliana. Exact buffer components were: 6% glycerol, 2% SDS, 50mM Tris-HCl pH6.8, 0.004% Bromophenol blue and 1% β-ME. Samples were denatured at 100 °C for 5 min, cooled down on ice, and were separated on 10% SDS-PAGE and blotted for 30 min/RT to PVDF (pore size of 0.2um), using semi-dry transfer. Blot was blocked with 5% milk 1 h/RT. Blot was incubated in the primary antibody at a dilution of 1: 1000 ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1: 10000 in 5% milk for 1h/RT with agitation. The blot was washed as above and developed with a following chemiluminescent detection reagent. Exposure tiume was 30 seconds.

Note: Background signal can be decreased by blocking ON/4°C and incubation of anti-FER antibodies 1h/RT

Courtesy of Dr. Jinggeng Zhou, Shanghai Normal University, China

Alternative name: Protein SIRENE