Lhcb3 | LHCII type III chlorophyll a/b-binding protein

Product no: AS01 002

AS01 002  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: Photosynthetic eukaryotes including A. thaliana, A. hypogaea, Ch. vulgaris, H. vulgare, L. esculentum (Solanum lycopersicon), M. crystallinum, N. tabacum, O. sativa, P. sativum, P. patens, Prasinoderma sp., Pyramimonas sp., P. vulgaris, S. oleracea, T. aestivum, Triticale, Z. mays

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  • Product Info
  • Immunogen:

    BSA-conjugated synthetic peptide derived from a highly conserved sequence of Lhcb3 proteins from angiosperms (monocots and dicots) and gymnosperms, including Arabidopsis thaliana Lhcb3 UniProt: Q9S7M0,TAIR:AT5G54270. This sequence is highly conserved even in Ginko biloba and one of the major LHCII-forms of Physcomitrella patens.

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Immunogen affinity purified serum in PBS pH 7.4
    Format: Lyophilized
    Quantity: 50 µg
    Reconstitution: For reconstitution add 50 µl of sterile water
    Storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
    Tested applications: Western blot (WB)
    Recommended dilution: 1 : 2000 (WB)
    Expected | apparent MW:

    28.7 | 26 kDa for Arabidopsis thaliana

  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana, Arachis hypogaea, Chlorella vulgaris, Cucumis sativa, Dactylis glomeRata, Hordeum vulgare, Lycopersicon esculentum (Solanum lycopersicon), Mesembryanthemum crystallinum, Nicotiana tabacum, Oryza sativa, Pisum sativum, Phaseolus vulgaris, Physcomirella patens, Prasinoderma sp., Pyramimonas sp., Spinacia oleracea, Triticum aestivum, Triticale, Zea may, Verbascum lychnitis
    Predicted reactivity:

    Cucumis melo, Dicots, Gymnosperms, Mosses

    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Western blot using anti-Lhcb3 antibodies

    1 - 10 ug of 4 days old of wild-type Arabidopsis thaliana seedlings extract

    2 - 5 ug of 4 days old of wild-type Arabidopsis thaliana seedlings extract

    3- 10 ug of 4 days old of gun5-1 mutant seedlings extract

    10 µg/well of total protein extracted from fresh 4 days old of Arabidopsis thaliana whole seedling.  Exact buffer components were: (50 mM Tris–HCl pH 7.5, 10% glycerol, 150 mM NaCl, 10 mM MgCl2, 5 mM EDTA, 5 mM DTT, 0.5% (v/v) Triton X-100, and 1 × protease inhibitors) and denatured with 4X SDS sample loading buffer (200 mM Tris-HCl (pH 6.8). 8% SDS (sodium dodecyl sulfate). 0.4% Bromophenol blue. 40% glycerol) at 95°C 10 min.  Samples were separated in the RT on 15 % SDS-PAGE and blotted for 0.5 h to PVDF (pore size of 0.2 um), using:  semi-dry at room temperature. Blot was blocked with 5 % milk for: 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:  2 000 at 4°C with agitation overnight. 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 RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated AS09 602 Agrisera) diluted to 1: 50 000 for 2 h/RT with agitation. The blot was washed as above and developed with a following chemiluminescent detection reagent: AS16 ECL-N-10 AgriseraBright (mid picogram). Exposure time was 5 seconds.

    Courtesy of Dr .Duorong Xu, LMU München, Germany Western blot using anti/Lhcb3 antibodies

    From 1 μg to 8 μg of chlorophyll from Arabidopsis thaliana chloroplasts extracted with 0.4 M sorbitol, 50 mM Hepes NaOH, pH 7.8, 10 mM NaCl, 5 mM MgCl 2 and 2 mM EDTA were loaded to lanes. Samples were denatured with Laemmli buffer at 75 0 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% milk for 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody Anti-Lhcb3 (LOT 1901) at a dilution of 1: 2000 in 1% milk in TBS-T overnight at 4 0 C with agitation. 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 HRP conjugated, from Agrisera, AS09 602) diluted to 1:20 000 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% H 2O2 in 0.1 M Tris- HCl, pH 8.5. Exposure time in ChemiDoc System was 240 seconds.

    Courtesy of Dr. Wioleta Wasilewska-Dębowska, University of Warsaw, Poland

    Application examples:

    Reactant: Mus musculus (House mouse)

    Application: Western Blotting

    Pudmed ID: 31245706

    Journal: Plant Direct

    Figure Number: 5B

    Published Date: 2018-02-01

    First Author: Rantala, S. & Tikkanen, M.

    Impact Factor: None

    Open Publication

    Stepwise detachment of photosynthetic protein complexes from the thylakoid membrane of WT, stn7, stn8, stn7stn8, and tap38/pph1. Thylakoid membranes of WT, stn7, stn8, stn7stn8, and tap38/pph1 plants harvested from moderate growth light (GL, 120 ?mol photons m?2 s?1) or after high light illumination (HL, 600 ?mol photons m?2 s?1 for 2 hr) were isolated and solubilized with 0%, 0.25%, 0.5%, 1%, and 2% DIG. Equal volumes of the soluble supernatant (S) and insoluble pellet (P) fractions were loaded and separated in SDS?PAGE followed by immunodetection of (a) proteins D1, PSAB, CYTF, and ATPF representing the protein complexes PSII, PSI, LHCII, Cyt b6f, and ATP synthase, respectively, as well as (b) proteins LHCB1, P?LHCB1, LHCB2, P?LHCB2, and LHCB3, representing the different subunits of the LHCII complexes. The differences in mutants with respect to WT as well as the differences in WT in response to Hl are marked with white boxes. Representative data from three different biological replicates are shown

  • Additional Information
  • Additional information:

    Antibody format is a total IgG fraction, which means that it is a pool of polyclonal antibodies obtained by purification of serum on Protein G, not on a specific antigen column.

    Additional information (application): Protein is processed into mature form (Jansson 1999).
  • Background
  • Background:

    The major light-harvesting antenna complex II (LHCII) in photsynthetic 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. While Lhcb1 and Lhcb2 are quite similar and regularily present in multiple gene-copies, the Lhcb3 protein differs in pigment-composition and molecular size and often is coded by only a single gene. Lhcb3 seems not to be present in the mobile LHCII trimers involved in state 1-state 2 transitions.
    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.

  • Product Citations
  • Selected references: von Bismarck et al. (2021) Light acclimation interacts with thylakoid ion transport to govern the dynamics of photosynthesis. Research Square; 2021. DOI: 10.21203/rs.3.rs-948381/v1.
    Wu et al. (2021). Formation of light-harvesting complex (LHC) II aggregates from LHCII-PSI-LHCI complexes in rice plants under high light. J Exp Bot. 2021 May 3:erab188. doi: 10.1093/jxb/erab188. Epub ahead of print. PMID: 33939808.
    Wojtowicz et al. (2020). Compensation Mechanism of the Photosynthetic Apparatus in Arabidopsis thaliana ch1 Mutants. Int J Mol Sci. 2020 Dec 28;22(1):221. doi: 10.3390/ijms22010221. PMID: 33379339; PMCID: PMC7794896.
    Koh et al. (2019). Heterologous synthesis of chlorophyll b in Nannochloropsis salina enhances growth and lipid production by increasing photosynthetic efficiency. Biotechnol Biofuels. 2019 May 14;12:122. doi: 10.1186/s13068-019-1462-3. eCollection 2019.
    Furukawa et al. (2019). Formation of a PSI–PSII megacomplex containing LHCSR and PsbS in the moss Physcomitrella patens. J Plant Res https://doi.org/10.1007/s10265-019-01138-2.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Oxygenic photosynthesis poster by prof. Govindjee and Dr. Shevela

    Z-scheme of photosynthetic electron transport by prof. Govindjee and Dr. Björn and Dr. Shevela

  • Reviews:
  • Reviews
    Below, you can grade the product on a scale from 0 to 5.
    Please also provide information about species, application, dilution and obtained result for the reviewed antibody.
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    Number of reviews: (4)
    Soo Yeon Ko | 2020-11-18
    0 | 0
    We always use this antibody when we check Lhcb3 band in Western blot(1:7000 dilution). It is working on Oriza Sativa.
    Soo Yeon Ko | 2019-11-18
    0 | 0
    The antibody is working on Oriza Sativa. We used isolated 2ug thylakoid membrane and can get a band (1:5000 dilution) clearly
    K. Browning | 2015-07-21
    0 | 0
    Works well with Arabidopsis total and chloroplast extracts. Some cross reactivity in total extracts with bands ~38 and ~75 kDa with overnight incubation in cold, but no cross reactive bands for chloroplast extracts when incubated only 1-2 hr at RT.
    Maciej Garstka | 2009-03-19
    0 | 0
    specific to pea, bean and rye


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