PsaD | PSI-D subunit of photosystem I
AS09 461 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Plants (monocots and dicots, conifers), moss: Physcomitrella patens, Chlamydomonas reinhardtii, Triticum aestivum, cyanobacteria
compartment marker of thylakoid membrane
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KLH-conjugated synthetic peptide 100% conserved in all known plant PsaD sequences including Arabidopsis thaliana PSI-D1 UniProt:Q9S7H1 , TAIR: At4g02770 and PSI-D2 UniProt: Q9SA56 , TAIR At1g03130 as well as Physcomitrella patens. The conservation in Chlamydomonas reinhardtii is high (14 of 16 aminoacids are identical).
17.9 | 20 (for Arabidopsis thaliana)
Alge, Dicots, Catalpa bungei, Cucumis melo, Conifers, Cyanidioschyzon merolae, Bigelowiella natans, Nannochloropsis sp. ,Nicotiana tabacum, Phaeodactylum tricornutum, Phyla dulcis, Zosteria marina
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Synechococcus elongatus sp. PCC 7942
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).
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
PsaD has frequently been used as a marker for intact PSI reaction centers.
This product can be sold containing proclin if requested.
Contains 0.1% ProClin.
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.
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