Lhca1 | PSI type I chlorophyll a/b-binding protein
AS01 005 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Monocots and dicots including A. thaliana, A. hypogaea, C. quitensis Kunt Bartl, E. crus-galli, F. margarita Swingle, G. hybrid, H. vulgare, L. esculentum (Solanum lycopersicum), N. tabacum, O. sativa, P. miliaceum, P. patens, P. sativum, P. strobus, P.vulgaris, S. oleracea, Z. mays
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BSA-conjugated synthetic peptide derived from the Lhca1 protein of Arabidopsis thaliana UniProt: Q01667, TAIR: At3g54890. This sequence is highly conserved in Lhca1 proteins of angiosperms (monocots and dicots) and gymnosperms.
25.99 | 22 kDa for Arabidopsis thaliana
1.0 µg of chlorophyll from mesophyll (M) and bundle sheath (BS) thylakoids of various C4 plants (Echinochloa crus-galli, Panicum miliaceum, 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 0C for 5 min and were separated on 12% SDS-PAGE and blotted 30 min to PVDF using wet transfer. Blot was blocked with 5% milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2000 overnight at 4°C with agitation in 1% milk 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, Lot 1702) 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 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 15 seconds.
Courtesy of Dr. Wioleta Wasilewska, Warsaw University, Poland
The light-harvesting protein Lhca1 is one of the four main and highly conserved types of chlorophyll a/b-binding proteins (Lhca1-4) of the light harvesting antenna (LHCI) of plant photosystem I. Lhca1 is imported as a precursor from the cytosol into the chloroplast. Upon insertion into the thylakoid membrane Lhca1 forms a heterodimer (LHCI-730) with Lhca4 that associates with the PSI core close to PsaG and PsaF.
A biochemical characterization of the plant LHCI antenna can be found in Klimmek et al. (2005) The structure of the higher plant light harvesting complex I: in vivo characterization and structural interdependence of the Lhca proteins. Biochemistry 44: 3065–3073
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