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product information
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| background |
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PsaA is a core protein of photosystem I. In plants and cyanobacteria, the primary step in oxygenic photosynthesis, the light induced charge separation, is driven bytwo large membrane intrinsic protein complexes, the photosystems I and II. Synonym: Photosystem I P700 chlorophyll a apoprotein A1. |
| immunogen |
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N-terminal part of recombinanat PsaA protein from Chlamydomonas reinhardtii P12154 |
| antibody format |
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rabbit |
polyclonal |
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serum |
lyophilized |
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| quantity |
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200 µl |
for reconstitution add 200 µl of sterile water. |
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| storage |
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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 |
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western blot (WB), immunogold (IG) |
| related products |
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collection of antibodies to PSI proteins recommended secondary antibody |
| additional information |
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PsaA is a hydrophobic protein and we recommend to use PVDF membrane for transfer to assure best results |
application information
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| recommended dilution |
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1: 1000 with standard ECL (WB), 1: 20 (IG) |
| expected | apparent MW |
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82 | 55-60 kDa |
| confirmed reactivity |
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Arabidopsis thaliana, Hordeum vulgare, Pisum sativum, Phaseolus vulgaris, Spinacia oleracea, Chlamydomonas reinhardtii, Synechococcus PCC 7942, Synechocystis PCC 6803 |
| predicted reactivity |
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dicots including: Lycopersicum esculentum, monocots including: Oryza sativa, Triticum aestivum trees: Picea spinulosa, Pinus thunbergii, Populus alba, Citrus x limon moss: Physcomitrella patens, algae, cyanobacteria |
| not reactive in |
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no confirmed exceptions from predicted reactivity known in the moment |
| additional information |
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immunogold localization has been done in leaf material of Arabidopsis thaliana |
| selected references |
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Rudowska et al. (2012). Chloroplast biogenesis - correlation between structure and function. BBA, available on line, March 2012. Pollari et al. (2011). Effects of deficiency and overdose of group 2 sigma factors in triple inactivation strains of Synechocystis sp. strain PCC 6803. J Bacteriol. 193(1):265-273. Fristedt et al. (2009). Phosphorylation of photosystem II controls functional macroscopic folding of photosynthetic membranes in Arabidopsis. Plant Cell 21:3950-3964
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application example
2 µg of total protein from (1) Arabidopsis thaliana leaf, (2) Hordeum vulgare leaf, (3) Chlamydomonas reinhardtii total cell, (4) Synechococcus sp. 7942 total cell all extracted with Protein Extration Buffer, 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 (GE Healthcare) 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, frecommended secondary antibody AS09 602) diluted to 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL Advance 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). Exposure time was 10 seconds.
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