PsbD | D2 protein of PSII (100 ĩl)
AS06 146-100 | Clonality: Polyclonal | Host: Rabbit | Reactivity: [global antibody] for A.thaliana, H. vulgare, L. corniculatus, N. tabacum, O. sativa, P. sativum, P. vulgaris, T. pRatense, U. prolifera, C. reinhardtii, Synechococcus sp. PCC 7942, Anabaena 7120, diatoms
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KLH-comjugated synthetic peptide derived from the C-terminal of known PsbD sequences including Arabidopsis pumila A4QJS8 , Hordeum vulgare P11849, Chlamydomonas reinhardtii P06007, Synechococcus sp. PCC 7002 P20898
39.4 | 28-30 kDa
Glycine max, Panax ginseng, Pinus thunbergii, Physcomitrella patens, Populus trichocarp, Solanum tuberosum, Spinacia oleracea, Triticum aestivum, Vitis vinifera, Zea mays
2 µg of total protein from (1) Arabidopsis thaliana leaf extracted with Protein Extraction 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, (5) Anabaena sp. 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% ECL Advance blocking reagent (GE Healthcare) 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, recommended secondary antibody AS09 602) diluted to 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL Advance 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). Exposure time was 3 seconds.
The peptide used to elicit this antibody has a perfect conservation across all full-length PsbD sequences from higher plants, lower
plants, cyanobacteria and unicellular algae except: -minor substitutions in some Prochlorococcus & Dinoflagellate sequences. The antibody should still work against these taxa, but it has not been tested yet. This antibody does not detect PsbA protein (D1).
There is a confirmed cross-reaction with TLA1 protein in Chlamydomonas reinhardtii.
For samples with a very low PSII content theremight be detection problems independent of the antibody. PSII proteins can vary in level depending upon liquid culture conditions. When the cells are in a stationary phase PSII content can drop to a very low level.
D2 protein (PsbD) forms the reaction core of PSII (Photosystem II) as a heterodimer with the D1 protein (PsbA). PsbD is homologous to the D1 protein, with slightly higher molecular mass of about 39,5 kDa. Accumulation of D2 protein is an important step in the assemply of the PSII reaction centre complex.
Huang et al. (2015). Rubisco accumulation is important for the greening of the fln2-4 mutant in Arabidopsis. Volume 236, July 2015, Pages 185–194.
Hojka et al. (2014). Inducible repression of nuclear-encoded subunits of the cytochrome b6f complex in tobacco reveals an extraordinarily long lifetime of the complex. Plant Physiol. 2014 Jun 24. pii: pp.114.243741.
Malnoë et al. (2014). Thylakoid FtsH Protease Contributes to Photosystem II and Cytochrome b6f Remodeling in Chlamydomonas reinhardtii under Stress Conditions. Plant Cell, Jan 21.