PsaK | PSI-K subunit of photosystem I

fusion protein between DHFR and the mature part of PsaK from Arabidopsis thaliana, UniProt: Q9SUI5, TAIR: At1g30380

Chlamydomonas reinhardtii, Cyanidioschyzon merolae strain 10D, Synechococcus sp. PCC 7942

Application example

2 µg of total protein from Arabidopsis thaliana leaf (1), Horderum vulgare leaf (2), Chlamydomonas reinhardtii total cell (3), Synechococcus sp. PCC 7942 total cell (4), all extracted with PEB (AS08 300) and separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% blocking reagent in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 10 000 for 1h at room temperature 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 room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:20 000 in 2% blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with chemiluminescent detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).

PsaK is a subunit of photosystem I. It has a role in organizing the peripheral light-harvesting complexes on the core antenna of photosystem I. Alternative names: photosystem I subunit X, PSI-K