Lhcb2 | LHCII type II chlorophyll a/b-binding protein
AS01 003 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Photosynthetic eukaryotes including A. thaliana, A. hypogaea, B. sylvaticum, C. quitensis Kunt Bartl, C. sativa, H. vulgare, C. reinhardtii, L. esculentum (Solanum lycopersicon), M. crystallinum, N. tabacum, O. sativa, P. patens, P. sativum, P. vulgaris, S. alba, S. oleracea, T. aestivum, Triticale, Z. mays
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BSA-conjugated synthetic peptide derived from a highly conserved sequence of Lhcb2 proteins from angiosperms (monocots and dicots) and gymnosperms, including Arabidopsis thaliana Lhcb2.1 UniProt: Q9SHR7, TAIR: AT2G05100, Lhcb2.2 UniProt: Q9S7J7, TAIR:AT2G05070, Lhcb2.3 UniProt:Q9XF87, TAIR:AT3G27690
28.6 | 25 kDa for Arabidopsis thaliana
Species and variants: Pea – Pisum sativum L. Bean – Phaseolus coccineus L. 3h – 3 hours of cold exposure 9h – 9 hours of cold exposure 12h – 12 hours of cold exposure 2d – 2 days of cold exposure
Samples of isolated thylakoids containing 3 µg of chlorophyll were denatured with Laemmli buffer (1 vol : 1 vol) at 75 °C for 5 min. Denatured samples containing 1 µg of chlorophyll were loaded in the gel wells, separated on 12% SDS-PAGE gels and blotted for 45 min at 100 V to PVDF membrane using wet transfer. Blot was blocked with 5% milk in TBS-T for 60 min at room temperature (RT) with agitation. The blot was incubated with the primary antibody at a dilution of 1:500 in 1% Amersham™ ECL Prime Blocking Agent in TBS-T overnight at 4ºC with agitation. The antibody solution was decanted and the blot was washed 3 times for 5 min in TBS-T at RT with agitation. The blot was incubated using a secondary antibody (goat anti-rabbit IgG HRP conjugated, from Agrisera, AS09 602) diluted to 1: 25 000 in 1% milk in TBS-T for 1h at RT with agitation. The blot was washed 5 times for 5 min in TBS-T, 1 time for 5 min in TBS, 1 time for 5 min in 0.1 M Tris (pH 8.5), and developed for 4 min in substrates (0.188 mM coumaric acid, 1.25 mM luminol, 0.01% H2O2). Exposure time was 5 seconds in ChemiDoc scanner (BioRad).
Msc Małgorzata Krysiak, Faculty of Biology, University of Warsaw, Poland
Immunoprecipitation has been done using Immunoprecipitation kit from Roche, Cat.No. 11 719 386 001.
Protein is processed into mature form (Jansson 1999).
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.
A molecular characterisation of the LHCII proteins can be found in Caffarri et al. (2004) A Look within LHCII: Differential Analysis of the Lhcb1−3 Complexes Building the Major Trimeric Antenna Complex of Higher-Plant Photosynthesis. Biochemistry 43 (29): 9467–9476.
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Hertle et al. (2020) A Sec14 Domain Protein Is Required for Photoautotrophic Growth and Chloroplast Vesicle Formation in Arabidopsis thaliana. Proc Natl Acad Sci USA 2020 Apr 3 (Immunogold)
Bethmann et al. (2019). The zeaxanthin epoxidase is degraded along with the D1 protein during photoinhibition of photosystem II. Plant Direct. 2019 Dec 1;3(11):e00185. doi: 10.1002/pld3.185.
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Rogowski et al. (2019). Photosynthesis and organization of maize mesophyll and bundle sheath thylakoids of plants grown in various light intensities. Environmental and Experimental Botany Volume 162, June 2019, Pages 72-86.
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Tadini et al. (2018). Trans-splicing of plastid rps12 transcripts, mediated by AtPPR4, is essential for embryo patterning in Arabidopsis thaliana. Planta. 2018 Jul;248(1):257-265. doi: 10.1007/s00425-018-2896-8.
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Shanmugabalaji et al. (2018). Chloroplast Biogenesis Controlled by DELLA-TOC159 Interaction in Early Plant Development. Curr Biol. 2018 Aug 20;28(16):2616-2623.e5. doi: 10.1016/j.cub.2018.06.006.
Du et al. (2018). Galactoglycerolipid Lipase PGD1 Is Involved in Thylakoid Membrane Remodeling in Response to Adverse Environmental Conditions in Chlamydomonas. Plant Cell. 2018 Feb;30(2):447-465. doi: 10.1105/tpc.17.00446.
Myouga et al. (2018). Stable accumulation of photosystem II requires ONE-HELIX PROTEIN1 (OHP1) of the light harvesting-like family. Plant Physiol. 2018 Feb 1. pii: pp.01782.2017. doi: 10.1104/pp.17.01782.
Kim et al. (2018). The rice zebra3 (z3) mutation disrupts citrate distribution and produces transverse dark-green/green variegation in mature leaves. Rice (N Y). 2018 Jan 5;11(1):1. doi: 10.1186/s12284-017-0196-8.
Rantala et al. (2017). Proteomic characterization of hierarchical megacomplex formation in Arabidopsis thylakoid membrane. Plant J. 2017 Dec;92(5):951-962. doi: 10.1111/tpj.13732.
Shin et al. (2017), Complementation of a mutation in CpSRP43 causing partial truncation of light-harvesting chlorophyll antenna in Chlorella vulgaris. Sci Rep. 2017 Dec 20;7(1):17929. doi: 10.1038/s41598-017-18221-0.
Cantrell and Peers (2017). A mutant of Chlamydomonas without LHCSR maintains high rates of photosynthesis, but has reduced cell division rates in sinusoidal light conditions. PLoS One. 2017 Jun 23;12(6):e0179395. doi: 10.1371/journal.pone.0179395.
Tyuereva et al. (2017). The absence of chlorophyll b affects lateral mobility of photosynthetic complexes and lipids in grana membranes of Arabidopsis and barley chlorina mutants. Photosynth Res. 2017 Apr 5. doi: 10.1007/s11120-017-0376-9. (Hordeum vulgare, western blot)
Míguez et al. (2017). Diversity of winter photoinhibitory responses: A case study in co-occurring lichens, mosses, herbs and woody plants from subalpine environments. Physiol Plant. 2017 Feb 14. doi: 10.1111/ppl.12551.
Yang-Er Chen et al. (2017). Responses of photosystem II and antioxidative systems to high light and high temperature co-stress in wheat. J. of Exp. Botany, Volume 135, March 2017, Pages 45–55.
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