PsaG | PSI-G subunit of photosystem I

4 µg/well of total chlorophyll from frozen thylakoids extracted freshly from Arabidopsis thaliana as described in Bassi et al., 1985 were separated on 4-15 % SDS-PAGE and blotted 1h to PVDF semi-dry. Blot was blocked with 5% milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 in TBS-T 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 Agrisera matching secondary antibody (anti-chicken IgG horse radish peroxidase conjugated AS010 1489) diluted to 1:5 000 in  for 1h/RT with agitation. The blot was washed as above and developed for 1 min with Agrisera ECLBright. Exposure time was 2 seconds.

Courtesy of Dr. Guillem Borràs I Gas, University of Copenhagen, Danmark