Lhcb1-P | LHCII type I chlorophyll a/b-binding protein, phopshorylated
AS13 2704 | Clonality: Polyclonal | Host: Rabbit | Reactivity:Arabidopsis thaliana
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KLH-conjugated synthetic peptide RKT*VAKPKGP, where T* indicates phospho-Thr
25 | 25 kDa for Arabidopsis thaliana
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1 ug of thylakoid membranes isolated from Arabidopsis thaliana wilde-type and 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, at a dilution of 1: 5 000/2.5 h, RT incubation, 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
The major light-harvesting antenna complex II (LHCII) in photosynthetic eukaryotes is located in the thylakoid membrane of the chloroplast. It is a heterotrimeric complex formed by up to 3 different individual subtypes of chlorophyll a/b-binding proteins: Lhcb1, Lhcb2, and Lhcb3. Lhcb1 is the most abundant chlorophyll a/b-binding protein in eukaryotic phototrophs and often is coded by several nuclear genes.
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