Anti-PsbS | 22 kDa Lhc-like PSII protein (chicken)

Chlamydomonas reinhardtii, Chlorella sp.

Application example

15 µg of Arabidopsis thaliana thylakoids from (1) PsbS-overexpressing line, (2) PsbS-deficient npq4-line, and (3) wt (col) together with (4) total leaf protein from Arabidopsis thaliana wt extracted with PEB (AS08 300) were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to nitrocellulose. Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-PsbS (AS03 032, 1:2000, 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 chemiluminescent reagent, using a Fuji LAS-3000 CCD (300s, standard sensitivity). Technical note(s): (a) This IgY shows reactivity to both markers loaded in lane M (MagicMark and NovexSharp, both Invitrogen), for comparison a marker lane from same filter (probed seperately with a IgG-antiserum) is shown to the right.  (b) the IgY reacts with 2 bands in leafs and thylakoids that both are absent in the PsbS-deficient npq4 line and therefore both might represent PsbS-forms with a difference in molecular weight of ~1 kDa.

Note: For this antibody, blocking needs to be increased to 10 % non-fat milk 1h/RT incubation.

The 22 kDa PsbS protein of photosystem II functions in the regulation of photosynthetic light harvesting. Along with a low thylakoid lumen pH and the presence of de-epoxidized xanthophylls, PsbS is necessary for photoprotective thermal dissipation of excess absorbed light energy in plants, measured as non-photochemical quenching of chlorophyll fluorescence.