Lhcb2-P | LHCII type II chlorophyll a/b-binding protein, phosphorylated
AS13 2705 | clonality: polyclonal | host: rabbit | reactivity:Arabidopsis thaliana, Echinochloa crus-galli, Zea mays
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1 : 10 000 (WB)
|Expected | apparent MW||
25 | 25 kDa for Arabidopsis thaliana
|Confirmed reactivity||Arabidopsis thaliana, Echinochloa crus-galli, Zea mays
|Predicted reactivity||Arachis hypogaea, Colobanthus quitensis Kunt Bartl, Hordeum vulgare, Lycopersicon esculentum (Solanum lycopersicon), Mesembryanthemum crystallinum, Nicotiana tabacum, Oryza sativa, Pisum sativum, Phaseolus vulgaris, Spinacia oleracea, Physcomitrella patens,|
|Not reactive in||
no confirmed exceptions from predicted reactivity are currently known
|Selected references||Fristedt et al. (2017). PSB33 sustains photosystem II D1 protein under fluctuating light conditions. Journal of Experimental Botany doi:10.1093/jxb/erx218.
Sato et al. (2015). Chlorophyll b degradation by chlorophyll b reductase under high-light conditions. Photosynth Res. 2015 Apr 21.
Leoni et al. (2013). Very rapid phosphorylation kinetics suggest a unique role for Lhcb2 during state transitions in Arabidopsis. Plant J. Oct;76(2):236-46. doi: 10.1111/tpj.12297. Epub 2013 Aug 26.
1 ug of thylakoid membranes isolated from Arabidopsis thaliana wild-type and respective mutants were solubilized with 3X LB (6 M urea, 12% SDS, 30% glycerol, 100 mM DTT, 150 mM Tris pH7.0, 0.8% Comassie G-250). 1 µg of total chlorophyll was loaded and separated on 16% SDS-PAGE, and then blotted for 2 h onto nitrocellulose membrane. Blots were blocked with milk powder for 2 h and then incubated in the primary antibody solution (AS13 2705, 1: 5 000) for 2.5 h, which was then decanted and the blot was washed 3 times for 5 min in TBST. Membrane was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h, followed by washing steps as above. All the steps fallowing transfer were performed in room temperature (RT) with agitation. Membrane was developed for 5 min with ECL according to the manufacturer’s instructions and recorded using FujiFilm CCD camera with 30 s increment time for around 5 min.
Courtesy of a phd candidate Małgorzata Pietrzykowska, Umeå Plant Science Centre, Sweden
Courtesy phD candidate Wiola Wasilewska, Faculty of Biology, University of Warsaw, Poland
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