Lhcb1 | LHCII type I chlorophyll a/b-binding protein (100 ĩg)
AS01 004-100 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Photosynthetic eukaryotes including A. thaliana, A. hypogaea, C. quitensis Kunt Bartl, H. vulgare, L. esculentum (Solanum lycopersicon), M. crystallinum, N. tabacum, O. sativa, P. sativum, P. vulgaris, S. vulgaris, S. oleracea, Z. mays
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BSA-conjugated synthetic peptide derived from a highly conserved sequence of Lhb1 proteins from angiosperms (monocots and dicots) and gymnosperms, includingArabidopsis thaliana At1g29910 (Lhcb1.1), At1g29920 (Lhcb1.2), At1g29930 (Lhcb1.3, most expressed), At2g34430 (Lhcb1.4), and At2g34420 (Lhcb1.5)
25 | 25 kDa for Arabidopsis thaliana
Dictos, Gymnosperms, Mosses, Triticum aestivum, Vitis vinifera
10 µg of total protein from (1) Arabidopsis thaliana leaf, (2) Hordeum vulgare leaf, (3) Zea mays leaf, (4) Chlamydomonas reinhardtii total cell, (5) Spinacia oleracea total leaf, (6) Physcomitrella patens, (7) Solanum tuberosum total leaf, (8) Solanum esculentum total leaf, all extracted with Protein Extraction Buffer PEB (AS08 300) were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2-2.5 % RPN2125 (GE Healthcare) in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 10 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS09 602) diluted to 1:25 000 in TBS-T for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with TMA-6 (Lumigen) detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad). Exposure time was 2 minutes.
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.
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