PsbO | 33 kDa of the oxygen evolving complex (OEC) of PSII (anti-protein)
AS06 142-33 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A.thaliana, C. reinhardtii, C. sp. DT, F. arundinacea cv. Kord, H. pluvialis, H. spontaneum, H. vulgare, N. benthamiana, P. abies, P. banksiana, S. oleracea, S. tuberosum cultivar Taedong Valley, Synechococcus sp. PCC 7942 and PCC6803, T. salsuginea, T.aestivum
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Native purified 33 kDa protein from Spinacia oleracea UniProt: P12359
35 | 33 kDa
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), Anabaena sp. total cell (5), 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.
Total IgG fraction has been purified by 40% ammonium sulpgate precipitation followed by DEAE cellulose chromatography
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).
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.
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.
Głowacka 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.