PsbP | 23 kDa protein of the oxygen evolving complex (OEC) of PSII (anti-peptide)

Chlamydomonas reinhardtii, Synechococcus sp. PCC 7942, Zostera marina

Application example

2 µg of total protein from (1) Arabidopsis thaliana leaf extracted with Protein Extration Buffer, PEB (AS08 300), (2) Hordeum vulgare leaf extracted with PEB, (3) Chlamydomonas reinhardtii total cell extracted with PEB, (4) Synechococcus sp. 7942 total cell extracted with PEB, were 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: 50 000 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 1:10 000 in 2% blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min withchemiluminescent 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).

PsbP - 23 kDa extrinsic protein of photosystem II (PSII). Processing of the protein results in a protein with molecular mass of around 20 kDa. PsbP is required to optimize water splitting process in PSII, by probaböy by optimisation of calcium and Cl- levels. The protein is found in higher plants and algae but is not conserved in cyanobacteria. Synonymes:Oxygen-evolving enhancer protein 2-1, chloroplastic, OEE2, 23 kDa subunit of oxygen evolving system of photosystem II, OEC 23 kDa subunit, OEC23, 23 kDa thylakoid membrane protein