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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1: 1000 with ECL (WB)
|Expected | apparent MW||
Arabidopsis thaliana, Chlamydomonas reinhardtii, Dactylis glomerata, Horderum vulgare, Mesembryanthemum sp.,Spinacia oleracea, Synechococcus PCC 7942, Cyanophora paradoxa, Flaveria sp. , Heterosigma akashiwo, Thalassiosira pseudonan, Euglena gracilis, Micromonas pusilla, Chlamydomonas reinhardtii, Porphyra sp., Gonyaulax polyedra, Emiliania huxleyi, symbiotic dinoflagellates of Stylophora pistillata and Turbinaria reniformis; Phaeodactylum tricornutum, Thalassiosira pseudonana
|Predicted reactivity||Chromera velia, Glycine max, Nicotiana tabacum, Spinacia oleracea, Physcomitrella patens, algae and cyanobacteria, Nannochloropsis sp., Prochlorococcus sp. (surface and a deep water ecotype), Synechococcus PCC 8801, Thermosynechococcus elongatus (BP-1)|
|Not reactive in||
no confirmed exceptions from predicted reactivity known in the moment
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.
|Selected references||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.
|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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