PsbO | 33 kDa of the oxygen evolving complex (OEC) of PSII (anti-protein)

Product no: AS06 142-33

AS06 142-33  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: A.thaliana, C. reinhardtii, C. vulgaris, C. sp. DT, E. crus-galli, F. arundinacea cv. Kord, H. pluvialis, H. spontaneum, H. vulgare, N. benthamiana, P. abies, P. banksiana, P. sativum, P. miliaceum, S. oleracea, S. tuberosum cultivar Taedong Valley, Synechococcus sp. PCC 7942 and PCC6803, T. salsuginea,
T.aestivum

compartment marker of thylakoid lumen

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  • Product Info
  • Immunogen:

    Native purified 33 kDa protein from Spinacia oleracea UniProt: P12359

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Total IgG. Protein G purified in PBS pH 7.4.
    Format: Lyophilized
    Quantity: 50 µl
    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: Immunoprecipitation (IP), Western blot (WB)
    Recommended dilution: 1 : 2000-1 : 5000 (WB)
    Expected | apparent MW:

    35 | 33 kDa

  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana, Chlamydomonas reinhardtii, Chlorella sp. DT, Chlorella vulgaris, Echinochloa crus-galli, Eucalyptus grandis x Eucalyptus camaldulensis, Festuca arundinacea cv. Kord, Haematococcus pluvialis, Hordeum spontaneum, Hordeum vulgare, Nicotiana benthamiana, Oryza sativa, Panicum miliaceum, Picea abies, Pinus banksiana, Pisum sativum, Spinacia oleracea, Solanum tuberosum cultivar Taedong Valley, Synechococcus sp. PCC7002, sp. PCC 7942 and sp. PCC6803, Thellungiella salsuginea, Triticum aestivum, Zea mays
    Predicted reactivity: Glycine max, Nicotiana tabacum, Solanum lycopersicum, Populus trichocarpa, Zosteria marina

    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Application example


    Western blot using anti-PsbO antibodies


    1 µg of chlorophyll from Pisum sativum thylakoids and from mesophyll (M) and bundle sheath (BS) thylakoids of Zea mays, Echinochloa crus-galli, Panicum miliaceum extracted with 0.4 M sorbitol, 50 mM Hepes NaOH, pH 7.8, 10 mM NaCl, 5 mM MgCl2 and 2 mM EDTA were loaded to lanes. 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 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody AS06 142-33 (Lot 1902) at a dilution of 1: 2000 overnight at 40C 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 HRP conjugated, from Agrisera, AS09 602, Lot 1902, as recommended) 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% H2O2 in 0.1 M Tris- HCl, pH 8.5. Exposure time in ChemiDoc System was 100 seconds

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



    western blot using anti-PsbO antibodies

    2 µg of total protein from Arabidopsis thaliana leaf (1), Hordeum vulgare leaf (2), Chlamydomonas reinhardtii total cell (3)Synechococcus sp. 7942 total cell (4), all extracted with 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% ECL Advance blocking reagent 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) diluted to 1:50 000 in 2% blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with chemiluminescent 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).

    Note:
    Detection level in algal species can be improved by adjustment of western blot protocol. Please inquire.

    Application examples:

    Reactant: Nicotiana tabacum (Common tobacco)

    Application: Western Blotting

    Pudmed ID: 29511193

    Journal: Nat Commun

    Figure Number: 1C

    Published Date: 2018-03-06

    First Author: G?owacka, K., Kromdijk, J., et al.

    Impact Factor: 13.783

    Open Publication

    Photosynthesis and water-use efficiency in Nicotiana tabacum plants with modified PsbS levels. aPsbS mRNA levels normalized to actin and tubulin sampled from fully expanded leaves of psbs-4, PSBS-28, PSBS-43 and wild-type (WT) N. tabacum plants. b PsbS protein levels normalized to the large subunit of oxygen-evolving complex of photosystem II (PsbO), determined from densitometry on immunoblots. c Representative immunoblot for PsbS and PsbO. d Net CO2 fixation rate (An), e NPQ, f quinone A (QA) redox state, and g stomatal conductance (gs) as a function of incident light intensity in fully expanded leaves. h Linear correlation between QA redox state and gs. Broken lines indicate measurements at highest light intensity. i Linear correlation between PsbS protein levels and intrinsic water-use efficiency (An/gs) at light intensity above 600??mol?m?2?s?1. Asterisks/lines show significant differences from WT (black for silencing, red for overexpressing lines; Dunnett’s two-way test; ??=?0.05). Error bars indicate s.e.m. (n?=?from 6 to 10 biological replicates)

  • Additional Information
  • Additional information:

    Total IgG fraction has been purified by 40% ammonium sulpgate precipitation followed by DEAE cellulose chromatography

    This product can be sold containing ProClin if requested

    Additional information (application):

    This antibody can be used as a loading control for Chlamydomonas reinhardtii while it not so suitable for higher plants as accumulation of these proteins might drop to 12.5-25 % of the WT level in mutants defective for PSII core (Schult et al. 2007).

  • Background
  • Background:

    PSII reaction centre components are  generating the redox potential required to drive highly oxidizing water splitting reaction. Four Mn atoms are present on a lumenal surface and form the catalyctic site of the water-splitting reaction which is in close association with the 33 kDa (PsbO), 23 kDa (PsbP) and 17 kDa (PsbQ) extrinistic subunits of oxygen evolving complex OEC. A 33-kDa extrinsic protein is also termed the Mn-stabilizing protein (MSP), however recent evidences shown that it is C-terminal domain of PsbA (D1) protein which is involved in in the assembly and stabilization of the OEC.

  • Product Citations
  • Selected references: Cecchin et al (2021) LPA2 protein is involved in photosystem II assembly in Chlamydomonas reinhardtii. Plant J. 2021 Jul 4. doi: 10.1111/tpj.15405. Epub ahead of print. PMID: 34218480.
    Loudya et al. (2021) Cellular and transcriptomic analyses reveal two-staged chloroplast biogenesis underpinning photosynthesis build-up in the wheat leaf. Genome Biol. 2021 May 11;22(1):151. doi: 10.1186/s13059-021-02366-3. PMID: 33975629; PMCID: PMC8111775.
    Terentyev (2020: The Main Structural and Functional Characteristics of Photosystem-II-Enriched Membranes Isolated From Wild Type and cia3 Mutant Chlamydomonas reinhardtii. Life (Basel). 2020 May 14;10(5):E63. doi: 10.3390/life10050063..
    Tang el al. (2020). OsNSUN2-Mediated 5-Methylcytosine mRNA Modification Enhances Rice Adaptation to High Temperature. Dev Cell. 2020 May 4;53(3):272-286.e7. doi: 10.1016/j.devcel.2020.03.009.
    Smythers et al. (2019). Characterizing the effect of Poast on Chlorella vulgaris, a non-target organism. Chemosphere Volume 219, March 2019, Pages 704-712.
    Vojta and Fulgosi (2019). Topology of TROL protein in thylakoid membranes of Arabidopsis thaliana. Physiol Plant. 2019 Jan 20. doi: 10.1111/ppl.12927.
    Da-Wei Yang et al. (2018). Genetically engineered hydrogenases promote biophotocatalysis-mediated H 2 production in the green alga Chlorella sp. DT. Int J of Hydrogen Energy.
    Liu et al. (2018). Effects of PSII Manganese-Stabilizing Protein Succinylation on Photosynthesis in the Model Cyanobacterium Synechococcus sp. PCC 7002. Plant Cell Physiol. 2018 Jul 1;59(7):1466-1482. doi: 10.1093/pcp/pcy080.
    G?owacka et al. (2018). Photosystem II Subunit S overexpression increases the efficiency of water use in a field-grown crop. Nat Commun. 2018 Mar 6;9(1):868. doi: 10.1038/s41467-018-03231-x.
    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.
    Sultan et al. (2017). The Reverse Transcriptase/RNA Maturase Protein MatR Is Required for the Splicing of Various Group II Introns in Brassicaceae Mitochondria. Plant Cell. 2016 Nov;28(11):2805-2829.
    Gandini et al. (2017). The transporter SynPAM71 is located in the plasma membrane and thylakoids, and mediates manganese tolerance in Synechocystis PCC6803. New Phytol. 2017 Mar 20. doi: 10.1111/nph.14526.
    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.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts
  • 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: (8)
    Soujanya Kuntam | 2021-11-09
    0 | 0
    We used this antibody to detect PsbO in complete protein extracts of C. reinhardtii using western blotting. It worked well without much optimization needed.
    Amanda Smythers | 2020-12-03
    0 | 0
    I used this antibody to help confirm psbO degradation in PSII-enriched membranes from spinach. It was easy to use and worked perfectly without optimization and helped lead to our publication: doi: 10.1016/j.chemosphere.2018.12.050
    Katya Georgieva | 2019-06-24
    0 | 0
    I used the antibody to check its expression during desiccation of Haberlea rhodopensis. PsbO is working perfect (1:2000 dilution) on isolated thylakoids and total leaf proteins. Clear, single band is obtained with ECL development.
    Qing-Bo | 2018-11-23
    0 | 0
    I used the antibody to detect the accumulation of the corresponding
    in Arabidopsis thaliana mutant with a dilution 1: 1000. The antibody works very well.
    Jun Liu | 2014-01-14
    0 | 0
    6 g of total protein from Arabidopsis was separated on 12 % SDS-page Gel , blotted 1h to NC membrane. Blots were incubated in the primary antibody at a dilution of 1: 20 000 overnight, A clear and specific band above 30 kDa has always been detected by ECL .
    Jerzy Kruk | 2013-11-21
    0 | 0
    Analysis perfomed on Arabidopsis total leaf extract, 10% SDS gel, 3 ug chlorophyll per lane, AB dilution 1:2000, the product observed at ca. 33 kDa
    Mayank Anand | 2010-06-16
    0 | 0
    I am satisfied with results I got, usinf Agrisera antibody. I used a 1:15000 dilution for this antibody. I am checking its expression at different stages of potato tuberization, so I took proteins from the stolons, initial tuber, maturing tubers and mature tubers.
    Irma Roberts | 2009-05-19
    0 | 0
    I used at a dilution of 1:20000 plus 1:150000 for the secondary antibody against PsbO from spinach, developing the membrane with ECL advance solution from GE.

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