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Lhcb2-P | LHCII type II chlorophyll a/b-binding protein, phosphorylated

323 €

AS13 2705  | clonality: polyclonal  |  host: rabbit  |  reactivity:Arabidopsis thaliana, Echinochloa crus-galli, Zea mays

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AS13 2705

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product information
Background

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. Lhcb2 is often coded by several nuclear genes and is found together with Lhcb1 within the mobile LHCII trimers involved in state1-state2 transition.

Immunogen

KLH-conjugated synthetic peptide: RRT*VKSTPQS, where T* indicates phospho-Thr

Host Rabbit
Clonality Polyclonal
Clone
Purity Affinity purified serum
Format Lyophilized
Quantity 25 ĩg
Reconstitution For reconstitution add 25 ĩl of sterile water.
Storage

store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications western blot (WB)
Related products

AS01 003 | anti-Lhcb2 | LHCII type II chlorophyll a/b-binding protein, rabbit antibodies

AS13 2704 | anti-Lhcb1-P | LHCII type I chlorophyll a/b-binding protein, phopshorylated, rabbit antibodies

All anti-LHC antibodies

Antibodies to other proteins involved in photosynthesis

Plant protein extraction buffer

Secondary antibodies

Additional information
application information
Recommended dilution

1 : 10 000 with standard ECL (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

Additional information
Selected references 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.

application example

western blot using anti-phosphorylated Lhcb2 antibodies

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 (AS01 004, 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

western blot using anti-Lhcb2-P | LHCII type II chlorophyll a/b-binding protein, phosphorylated on maize samples

1.0 µg of chlorophyll from mesophyll (M) and bundle sheath (BS) thylakoids of various treatments of Zea mays extracted with 0.4 M sorbitol, 50 mM Hepes NaOH, pH 7.8, 10 mM NaCl, 5 mM MgCl2 and 2 mM EDTA. Samples were denatured with Laemmli buffer at 75°C for 5 min and were separated on 12% SDS-PAGE and blotted 30 min to PVDF using wet transfer. Blot was blocked with 5% BSA for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10 000 overnight at 4°C with agitation in 1% BSA in TBS-T. The antibody solution was decanted and the blot was washed 4 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:25 000 in  1 % BSA in TBS-T for 1h at RT with agitation. The blot was washed 5 times for 5 min in TBS-T and 2 times for 5 min in TBS, and developed for 1 min with 1.25 mM luminol, 0.198 mM coumaric acid and 0.009% H2O2 in 0.1 M Tris- HCl, pH 8.5. Exposure time in ChemiDoc System was 54 seconds.

Courtesy phD candidate Wiola Wasilewska, Faculty of Biology, University of Warsaw, Poland

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