Anti-PsbF | beta subunit of Cytochrome b559 of PSII

Chlamydomonas reinhardtii, cyanobacteria

Application example

5µg protein of Arabidopsis thaliana (1) leaf, extracted with PEB (AS08 300), (2) chloroplasts (normal light), (3) thylakoids (extracted from 2), (4) chloroplasts (high light), (5) thylakoids (extracted from 4), (6) Photosystem I, and (7) BBY particles (enriched in Photosystem II) from pea were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1.5h (30V) to nitrocellulose. Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with AS06 113 (1:1000, 1h) and secondary anti-rabbit (1:20000, 1 h) antibody (HRP conjugated) in TBS-T containing 2% low fat milk powder. Antibody incubations were followed by washings in TBS-T (15, +5, +5, +5 min). All steps were performed at RT with agitation. Signal was detected with chemiluminescent detection reagent, using a Fuji LAS-3000 CCD (150s, high sensitivity). The PsbF protein is detected with an apparent mass of < 10 kDa
With this protocol, there is a cross-reaction with a target at ~43 kDa.

Cytochrome b559 (Cyt b559) is encoded by the chloroplast genes psbE and psbF and is comprised of two low molecular mass polypeptides, α and ß subunits, with molecular masses of 9 and 4 kDa, respectively. The Cyt b559 is closely associated with PSII in all oxygenic photosynthetic organisms. The α and ß subunits of the Cyt b559 are components of the minimal PSII reaction center complex that is still capable of primary charge separation In summary, both PsbE and PsbF are essential components for PSII assembly, and they are probably involved in electron transport mechanisms that help to protect PSII from photodamage.