PsaD | PSI-D subunit of photosystem I

AS09 461 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Plants (monocots and dicots, conifers), moss: Physcomitrella patens, Chlamydomonas reinhardtii, cyanobacteria

Product Information

Immunogen

KLH-conjugated synthetic peptide 100% conserved in all known plant PsaD sequences including Arabidopsis thaliana (At1g03130 and At4g02770) as well as Physcomitrella patens. The conservation in Chlamydomonas reinhardtii is high (14 of 16 aminoacids are identical).

Host Rabbit

Clonality Polyclonal

Purity Serum

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 tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications Immunoprecipitation (IP), Western blot (WB)

Recommended dilution 1 : 1000 (WB)

Expected | apparent MW

17.9 | 20 (for Arabidopsis thaliana)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Chlamydomonas reinhardtii, Hordeum vulgare, Lactuca sativa, Oryza sativa, Physcomitrella patens, Picea glauca, Pinus strobus, Oryza sativa, Physcomitrella patens, Spinacia oleracea, Synechocystis PCC 6803, Triticum aestivum, Triticale, Zea mays

Predicted reactivity

Alge, Dicots, Catalpa bungei, Cucumis melo, Conifers, Cyanidioschyzon merolae, Bigelowiella natans, Nicotiana tabacum, Phaeodactylum tricornutum, Phyla dulcis, Zosteria marina

Not reactive in

Synechococcus elongatus sp. PCC 7942

Application examples

Application example

10 µg of total leaf protein extracted with PEB (AS08 300) from (1) Zea mays, (2) Chlamydomonas reinhardtii, and (3) Spinacia oleracea were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 80 min (30V) to nitrocellulose. Filter was blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-PsaD (AS09 461, 1:1000, 1h) and secondary anti-rabbit (1:40000, 1h) antibody (HRP conjugated) in TBS-T containing 2% low fat milk powder. Antibody incubations were followed by washings in TBS-T (15, +5, +5, +5 min). All steps were performed at RT with agitation. Signal was detected with chemiluminescent detection reagent using a GenoPlex Chemi CCD (accumulated signal 10 x 30s exposure, bin 2x2).

Western blot with anti-PsaD antibodies on Chlamydomonas reinhardtii cell extracts

Total cellular (lanes 2 – 5) and membrane proteins (lanes 6 – 9) from various environmental isolated of Chlamydomonas reinhardtii were extracted with a buffer containing 62.5mM Tris-HCl pH 6.8, 10% glycerol, 2% SDS, 50mM DTT, 10mM NaF and 1% protease inhibitors (P9599, Sigma Aldrich) and denatured at 65°C for 5 min. Samples (0.25 µg of chlorophyll per lane) were separated on 12% SDS-PAGE containing 6M urea and blotted 1h to PVDF using tank transfer. Blots were blocked with 5% skim milk powder in TBS-T for 1h at room temperature (RT) with agitation. Blots were incubated in the primary antibody at a dilution of 1:5000 overnight at 4°C. The antibody solution was decanted and the blots were rinsed briefly once, then washed 3 times for 10 min in TBS-T at RT with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG HRP-conjugated, Agrisera AS09 602) diluted to 1:20 000 for 1h at RT with agitation. The blots were washed as above, developed for 5 min with chemiluminescent detection reagent and then imaged using a ChemiDoc MP imaging system and Image Lab software (Bio-Rad Laboratories). Exposure time was 10 seconds.

Courtesy of Kenneth Wilson, University of Saskatchewan, Canada

Additional information

PsaD has frequently been used as a marker for intact PSI reaction centers.

This product can be sold containing proclin if requested.

This antibody is a replacement for former product, anti-PsaD AS04 046

Contains 0.1% ProClin.

Related products

PSI available antibodies to Photosystem I proteins

Photosynthesis available antibodies to photosynthetic proteins

Plant protein extraction buffer

Secondary antibodies

Background

PsaD (PSI-D) is a core subunit of photosystem I highly conserved in all photosynthetic organisms (including bacteria with Fe-S type reaction centers). In eukaryots its encoded by 1 to 2 nuclear gene(s) and imported as a precursor into the chloroplast. In the thylakoid membrane it associates with PsaA and PsaB on the stromal site of the PSI core forming the Fd-docking site. PsaD is also required for the stable assembly of PsaC.

Product citations

Selected references 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.
Storti et al. (2018). Role of cyclic and pseudo-cyclic electron transport in response to dynamic light changes in Physcomitrella patens. Plant Cell Environ. 2018 Nov 29. doi: 10.1111/pce.13493.
Mao et al. (2018). Comparison on Photosynthesis and Antioxidant Defense Systems in Wheat with Different Ploidy Levels and Octoploid Triticale. Int J Mol Sci. 2018 Oct 2;19(10). pii: E3006. doi: 10.3390/ijms19103006.
Nama et al. (2018). Non-photochemical quenching-dependent acclimation and thylakoid organization of Chlamydomonas reinhardtii to high light stress. Photosynth Res. 2018 Jul 7. doi: 10.1007/s11120-018-0551-7.
Gao et al. (2018). Effect of green light on the amount and activity of NDH-1–PSI supercomplex in Synechocystis sp. strain PCC 6803. Photosynthetica (2018) 56: 316. https://doi.org/10.1007/s11099-018-0790-z.
Kong et al. (2018) Interorganelle Communication: Peroxisomal MALATE DEHYDROGENASE2 Connects Lipid Catabolism to Photosynthesis through Redox Coupling in Chlamydomonas. Plant Cell. 2018 Aug;30(8):1824-1847. doi: 10.1105/tpc.18.00361
Li et al. (2018). Modulating plant growth-metabolism coordination for sustainable agriculture. Nature. 2018 Aug 15. doi: 10.1038/s41586-018-0415-5.
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.
Merry et al. (2017). A comparison of pine and spruce in recovery from winter stress; changes in recovery kinetics, and the abundance and phosphorylation status of photosynthetic proteins during winter. Tree Physiol. 2017 Sep 1;37(9):1239-1250. doi: 10.1093/treephys/tpx065.
Ge at al. (2017). Translating Divergent Environmental Stresses into a Common Proteome Response through the Histidine Kinase 33 (Hik33) in a Model Cyanobacterium. Mol Cell Proteomics. 2017 Jul;16(7):1258-1274. doi: 10.1074/mcp.M116.068080.
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.
Gerotto et al. (2016). Flavodiiron proteins act as safety valve for electrons in Physcomitrella patens. PNAS DOI 10.1073.
Heinnickel et al. (2016). Tetratricopeptide repeat protein protects photosystem I from oxidative disruption during assembly. Proc Natl Acad Sci U S A. 2016 Mar 8;113(10):2774-9. doi: 10.1073/pnas.1524040113
Fujii et al. (2015). Photoprotection vs Photoinhibition of Photosystem II in Transplastomic Lettuce (Lactuca sativa) Dominantly Accumulating Astaxanthin. Plant Cell Physiol. 2015 Dec 7. pii: pcv187.
Daddy et al. (2015). A novel high light-inducible carotenoid-binding protein complex in the thylakoid membranes of Synechocystis PCC 6803. Sci Rep. 2015 Mar 30;5:9480. doi: 10.1038/srep09480.
Qin et al. (2014). Isolation and characterization of a PSI-LHCI super-complex and its sub-complexes from a siphonaceous marine green alga, Bryopsis Corticulans. Photosynth Res. 2014 Sep 12.
Cheng and He (2014). PfsR Is a Key Regulator of Iron Homeostasis in Synechocystis PCC 6803. PLoS One. 2014 Jul 10;9(7):e101743. doi: 10.1371/journal.pone.0101743. eCollection 2014.
Tomizioli et al. (2014). Deciphering thylakoid sub-compartments using a mass spectrometry-based approach. Mol Cell Proteomics. 2014 May 28. pii: mcp.M114.040923.