Anti-PsaE | PSI-E subunit of photosystem I (affinity purified)

Application example

Thylakoid membranes (10 µg of total chlorophyll) extracted freshly from Hordeum vulgare leaves with 100 mM HEPES-KOH (pH 7.5), 0.3 M sorbitol, 2 mM EDTA, and 1mM MgCl2 and denatured with a Laemmli buffer at 80°C for 5 min were separated on 12% SDS-PAGE and blotted 1 h to nitrocellulose (pore size of 0.2 um), using semi-dry transfer. Blot was blocked with 4% milk for 2 h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:5000 (PsaE) for 1 h/RT with agitation in PBS-T. The antibody solution was decanted and the blot was rinsed briefly, then washed 3 times for 5 min in PBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:25000 in for 1 h/RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent according to manufacture's recommendations. Exposure time was 30 seconds.

Courtesy Dr. Anja Liszkay, CNRS, France

PsaE is a nucleus encoded subunit of the Photosystem I reaction center. It is located on the stroma side and interacts with PsaF. PsaE may be involved in Fd reduction.