PsaC | PSI-C core subunit of photosystem I

AS10 939 | Clonality: Polyclonal | Host: Rabbit | Reactivity: [global antibody] for higher plants, algae, cyanobacteria, diatoms

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product information
Background		PsaC is a conserved, chloroplast-encoded, Fe-S binding protein of approximately 10kDa, present in all known Photosystem I complexes. It is located on the stromal side of the thylacoid membranes. PsaC coordinates the Fe–S clusters FA and FB through two cysteine-rich domains.
Immunogen		KLH-conjugated synthetic peptide conserved in all known PsaC proteins including Arabidopsis thaliana AtCg01060, Hordeum vulgare P69416, Oryza sativa P0C360, Chlamydomonas reinhardtii Q00914, Synechococcus elongatus Q31QV2
Host		Rabbit
Clonality		Polyclonal
Clone
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		Western blot (WB)
Related products		AS10 939-10 \| PsaC \| PSI-C core subunit of photosystem I AS04 042P \| PsaC \| PSI-C core subunit of photosystem I (PsaC antibody + PsaC protein positive control) AS04 042S \| PsaC protein standard for quantitation or positive control collection of antibodies to PSI proteins Plant and algal protein extraction buffer Secondary antibodies
Additional information		Peptide target used to elicit this antibody is well conserved in all photoautotrophs except some cyanobacteria, some red algae and Cyanophora paradoxa, which contain a conserved substitution of a valine to an isoleucine. The performance of the antibodies has been confirmed against taxa containing both the valine and isoleucine variants. Example of a simulataneous western blot detection with RbcL, PsbA and PsaC antibodies. More information about quantitative western blot using PsaC antibody can be found here.

application information
Recommended dilution		1 : 1000 (WB)
Expected \| apparent MW		9 kDa
Confirmed reactivity		Arabidopsis thaliana, Chlamydomonas reinhardtii, Cyanophora paradoxa, Dactylis glomeRata, Emiliania huxleyi, Euglena gracilis, Gonyaulax polyedra, Horderum vulgare, Mesembryanthemum sp., Spinacia oleracea, Flaveria sp., Heterosigma akashiwo,Micromonas pusilla, Phaeodactylum tricornutum, Porphyra sp., symbiotic dinoflagellates of Stylophora pistillata and Turbinaria reniformis; Synechococcus PCC 7942, Synechocystis sp. PCC 6803, Thalassiosira pseudonana, Thalassiosira punctigera, Triticum aestivum
Predicted reactivity		Algae, Cannabis sativa, Chromera velia, Cyanobacteria, Glycine max, Nannochloropsis sp., Nicotiana tabacum, Physcomitrella patens, , Prochlorococcus sp. (surface and a deep water ecotype), Spinacia oleracea, Synechococcus PCC 8801, Thermosynechococcus elongatus (BP-1)
Not reactive in		No confirmed exceptions from predicted reactivity are currently known.
Additional information		In some species minor cross reactions with some larger proteins are seen. These may contain related iron-sulfur binding motifs. Therefore size verification of the reacting band is required. Due to the small size of the protein, care should be taken to differentiate between chemiluminescent signal from PsaC and non-specific signals from chlotophylls or lipids if pigment is retained near the bottom of the blot. For the most optimal results use:thylakoid membranes or PSI particles, solubilized in a SDS sample buffer (final concentrations: 63 mM Tris HCl, 10% glycerol, 2% SDS, 0.0025% bromophenol blue) with 2.5% beta-mercaptoethanol at 85C for 2 minutes. The samples were spun softly, then the supernatant loaded. This product can be sold containing ProClin if requested
Selected references		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. Cantrell and Peers (2017). A mutant of Chlamydomonas without LHCSR maintains high rates of photosynthesis, but has reduced cell division rates in sinusoidal light conditions. PLoS One. 2017 Jun 23;12(6):e0179395. doi: 10.1371/journal.pone.0179395. Zang et al. (2017). Characterization of the sulfur-formation (suf) genes in Synechocystis sp. PCC 6803 under photoautotrophic and heterotrophic growth conditions. Planta. 2017 Jul 14. doi: 10.1007/s00425-017-2738-0. Hu et al. (2017). The SUFBC2 D Complex is Required for the Biogenesis of All Major Classes of Plastid Fe-S Proteins. Plant J. 2017 Jan 19. doi: 10.1111/tpj.13483. 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. Li et al. (2016). A Hard Day's Night: Diatoms Continue Recycling Photosystem II in the Dark. Front. Mar. Sci., 08 November 2016 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 Rozpądek et al. (2015). The fungal endophyte Epichloë typhina improves photosynthesis efficiency of its host orchard grass (Dactylis glomerata). Planta. 2015 Jun 10. Subramanyam et al. (2014). Structural and functional changes of PSI-LHCI supercomplexes of Chlamydomonas reinhardtii cells grown under high salt conditions. Planta. 2010 Mar;231(4):913-22. Dang et al. (2014). Combined Increases in Mitochondrial Cooperation and Oxygen Photoreduction Compensate for Deficiency in Cyclic Electron Flow in Chlamydomonas reinhardtii. Plant Cell. 2014 Jul 2. pii: tpc.114.126375.

application example

5 µg of total protein from samples such as (1) Arabidopsis thaliana leaf, (2) Hordeum vulgare leaf, (3) Chlamydomonas reinhardtii total cell, (4) Synechococcus sp. 7942 total cell, (5) PsaC protein standard (AS04 042S), total protein from all the samples were extracted with Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70°C for 5 min and kept on ice before loading. Protein samples were separated on NuPAGE 4-12% Tris-Bis gel (Invitrogen) LDS-PAGE and blotted for 1h to 1.5h on PVDF using tank transfer. Blots were blocked immediately following transfer in 2% ECL Advance blocking reagent for 1h at RT with agitation. Blots were incubated with PsaC antibody at a dilution of 1: 10 000 (in blocking reagent) for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at RT with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG HRP conjugated, AS09 602) diluted to 1:50 000 in blocking reagent for 1h at RT with agitation. The blots were washed as above. The blot was developed for 5 min with ECL Advance (GE Healthcare). Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).

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