PsbTn | Tn protein of PSII

10 µg of protein from Arabidopsis thaliana thylakoids (Th), BBY, stroma (St), lumen (LU) were separated on  16.5 % SDS-PAGE (Tris-Tricine mini-gel containing 6 M urea) and blotted 1h to PVDF using semi-dry. Blot was blocked with 5 % milk for 1h/RT or 4°C/ON with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10 000 for 1h/RT with agitation in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 2x for 10 min with TBS and 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in a matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:50 000 for 1h/RT with agitation. The blot was washed as above and developed with chemiluminescent detection reagent. Exposure time was 3 seconds. 

Courtesy of Patrik Storm, Umeå University, Sweden