Lhcb4 | CP29 chlorophyll a/b binding protein of plant PSII

Product no: AS04 045

AS04 045  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: A. thaliana, C. sinensis, C. sativus, H. vulgare, N. tabacum, O. sativa, P. sativum, P. vulgaris, T. aestivum, Triticale, Z. mays

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  • Product Info
  • Immunogen:

    BSA-conjugated synthetic peptide derived from a highly conserved sequence of Lhb4 proteins from angiosperms (monocots and dicots) and gymnosperms, including Arabidopsis thaliana (Lhcb4.1 At5g01530 and Lhcb4.2 At3g08940 and Lhcb4.3 At2G40100).

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Total IgG. Protein G purified in PBS pH 7.4.
    Format: Lyophilized
    Quantity: 0.5 mg
    Reconstitution: For reconstitution add 250 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 : 7 000 (WB)
    Expected | apparent MW:

    31.9 | 29 kDa for Arabidopsis thaliana

  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana, Camelina sinensis, Cucumis sativus L. cv. Jihong no. 2, Drosera capensis, Hordeum vulgare, Lactuca sativa, Nicotiana tabacum, Oryza sativa, Pisum sativum, Phaseolus vulgaris, Triticum aestivum, Triticale, Zea mays
    Predicted reactivity:

    Catalpa bungei, Cucumis sativus, Malus domestica, Populus, gymnosperms and microalgae Ostrecococcus tauri; the target sequence is only weakly conserved in Physcomitrium patens


    Species of your interest not listed? Contact us
    Not reactive in:

    Chlamydomonas reinhardtii (please use AS06 117 for this organism)

  • Application Examples
  • Application example

    Western blot using anti-Lhcb4 antibodies

    5 µg of total protein from embebed seeds of Nicotiana tabacum growing during 4 d in dark (0) and then transfer to continue light growing for 6 h (6) extracted with LB2x buffer and denatured 90 ºC for 2-5 min, were separated on 12.5 % SDS-PAGE and blotted 1h to PVDF using tank transfer. Blots were blocked with TBS-T with 5% dry-milk for 3h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10 000 overnight at 4 ºC with agitation in TBS-T with 5% dry-milk. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 4 times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602, from Agrisera) diluted to 1:30 000 in TBS-T with 5% dry-milk for 1h at RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection. Exposure time was 60 seconds.

    Courtesy of Dr. Concha Almoguera, Inst. de Recursos Naturales y Agrobiología –CSIC, Spain
    Application examples:

    Reactant: Arabidopsis thaliana (Thale cress)

    Application: Western Blotting

    Pudmed ID: 25835989

    Journal: PLoS One

    Figure Number: 7A

    Published Date: 2015-04-04

    First Author: Fristedt, R., Martins, N. F., et al.

    Impact Factor: 2.942

    Open Publication

    Altered protein accumulation and stability of the chloroplast ATP synthase in the cgl160 mutant visualized by immunoblotting.A. Immunoblots with antibodies against essential subunits of the photosynthetic protein complexes of wild-type (Col-4) Arabidopsis and the two cgl160 T-DNA insertion lines grown under long-day and short-day conditions. Isolated thylakoid membranes were used, and equal amounts of chlorophyll were loaded onto the SDS-PAGE gel. For approximate quantification, wild-type samples from long-day plants were diluted to 10%, 25% and 50%, respectively. Accumulation of PSII was probed with antibodies against PsbB and PSBO. Additionally, the PSBS protein involved in NPQ and the minor PSII antenna protein LHCB4 were probed. Accumulation of the cytochrome b6f complex was probed with antibodies against the essential subunits PetA (cytochrome f), PetB (cytochrome b6), and PETC (Rieske protein). Accumulation of PSI was probed with antibodies against the reaction center subunit PsaB and the stromal ridge subunit PsaD. ATP synthase accumulation was probed with antibodies against the CF1 subunits AtpA (CF1?), AtpB (CF1?) and AtpD (CF1?) and antibodies against the CF0 subunits AtpF (CF0b) and AtpI (CF0a). B. Loading difference estimation for immunoblotting CF1 between wild type and cgl160-1. To obtain similar immunoblotting signal three times more (15 ?g protein) was needed for cgl160-1 compared to wild type (5 ?g protein). C. Maintenance of CF1 was measured by incubating leaves from wild type and cgl160-1 in solution containing the plastid protein synthesis inhibitor chloramphenicol for the indicated time points. Protein extract was isolated and separated by SDS-PAGE, immunoblotted and probed with specific antibodies against CF1 and LHCB2.1. Three times more protein was loaded from the mutant to obtain equal level of CF1 immunoblotting signal, as specified in B.


    Reactant: Arabidopsis thaliana (Thale cress)

    Application: Western Blotting

    Pudmed ID: 31249292

    Journal: Nat Commun

    Figure Number: 1A,B

    Published Date: 2019-06-27

    First Author: Dogra, V., Li, M., et al.

    Impact Factor: 13.783

    Open Publication

    Light- and 1O2-dependent EX1 degradation. Continuous light (CL)-grown 5-day-(d)-old transgenic seedlings of aex1 and bex1 flu expressing EX1-GFP under the control of the 35S promoter were transferred to the dark (for 2, 4, 8?h) and the 8-h-dark (D)-treated seedlings were re-exposed to light (for 0.5, 1, 2?h) at the light intensity of 100?µmolm-2?s-1. Total protein was extracted and analyzed by western blot. Chlorophyll a/b binding protein CP29 (Lhcb4) and cytosolic UDP-glucose pyrophosphorylase (UGP) were used as controls. EX1-GFP, Lhcb4, and UGPase were detected using antibodies against GFP, Lhcb4, and UGP, respectively. c The levels of EX1-GFP in the dark or after re-exposing to light were compared to its abundances under CL conditions. Average intensity values of the protein bands were calculated using AzureSpot software v14.0 (AZURE). Data represents the mean of three biological repeats. Error bars show standard error of the mean. Asterisks in c indicate statistically significant differences to CL condition (P?<?0.05, Student’s t-test)

  • Additional Information
  • Additional information:

    An overview about the different Lhc-protein types in plants can be found in Klimmek et al. (2006) Abundantly and rarely expressed Lhc protein genes exhibit distinct regulation patterns in plants. Plant Physiol 140: 793-804.

    Lhcb4 protein is processed into mature form (Jansson 1999).

  • Background
  • Background:

    Lhcb4 (CP29) is one of the 3 minor chlrorophyll a/b-binding proteins associated with Photosystem II in plants and algae. Lhcb4 has been suggested to act in the regulation of the chl a excited state concentration of PSII because of its ability of sensing lumenal pH resulting in reversible phosphorylation. In Arabidopsis thaliana 2 genes code for two isoforms Lhcb4.1 and Lhcb4.2. A third isoform (Lhcb4.3, At2g40100), probably only present in dicots, has found to be differently regulated and therefore has been denoted as Lhcb8.

  • Product Citations
  • Selected references: Ye et al. (2023). The light-harvesting chlorophyll a/b-binding proteins of photosystem II family members are responsible for temperature sensitivity and leaf color phenotype in albino tea plant. J Adv Res . 2023 Dec 25:S2090-1232(23)00404-6.doi: 10.1016/j.jare.2023.12.017.
    Hao and Malnoë (2023). A Simple Sonication Method to Isolate the Chloroplast Lumen in Arabidopsis thaliana.Bio Protoc. 2023 Aug 5; 13(15): e4756.  
    Cazzaniga et al. (2022). Engineering astaxanthin accumulation reduces photoinhibition and increases biomass productivity under high light in Chlamydomonas reinhardtii. Biotechnol Biofuels Bioprod. 2022 Jul 11;15(1):77. doi: 10.1186/s13068-022-02173-3. PMID: 35820961; PMCID: PMC9277849.
    Bru, Steen, Park, et al. (2022) The major trimeric antenna complexes serve as a site for qH-energy dissipation in plants. J Biol Chem. 2022;298(11):102519. doi:10.1016/j.jbc.2022.102523
    Pavlovic & Kocab. (2021) Alternative oxidase (AOX) in the carnivorous pitcher plants of the genus Nepenthes: what is it good for? Ann Bot. 2021 Dec 18:mcab151. doi: 10.1093/aob/mcab151. Epub ahead of print. PMID: 34922341.
    Fukura et al. (2021) Enrichment of chlorophyll catabolic enzymes in grana margins and their cooperation in catabolic reactions. J Plant Physiol. 2021 Nov;266:153535. doi: 10.1016/j.jplph.2021.153535. Epub 2021 Sep 25. PMID: 34607178.
    Chen et al. (2021)Degradation of the photosystem II core complex is independent of chlorophyll degradation mediated by Stay-Green Mg2+ dechelatase in Arabidopsis,Plant Science,Volume 307,2021,110902,ISSN 0168-9452,https://doi.org/10.1016/j.plantsci.2021.110902.
  • 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.
    Your name will be displayed as the sender.
    Number of reviews: (5)
    Kamil Trzebuniak | 2020-03-06
    0 | 0
    I have used this antibody in my research on Arabidopsis thaliana and it has worked well. For my Western Blott I have used the antibody with the recommended dilution (1:7000) while the total protein load was 20 ug per well. The obtained results were satisfying, the protein mass was about 30 kDA as expected.

    Kamil,
    Jagiellonian University
    I used this antibody in my reserch on Arabidopsis thaliana and it worked really well
    Soo Yeon Ko | 2019-11-18
    0 | 0
    The antibody is working on Oriza Sativa. We used isolated 2ug thylakoid membrane and can get CP29 band (1:10000 dilution) clearly
    Katya Georgieva | 2019-06-24
    0 | 0
    The antibody is working very well on Haberlea rhodopensis. We used isolated thylakoid proteins (3ug chlorophyll per lane) and CP29 (1:7000 dilution) gave clear single band.
    Monica Colombo | 2015-01-30
    0 | 0
    The antibody work with a high dilution 1:7000 in Arabidopsis thaliana with 3ug of proteins
    Shizue Matsubara | 2012-04-04
    0 | 0
    This antibody gave a clear single band with Arabidopsis leaf protein extracts.

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