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

Product Information

Immunogen

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).

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, Nannochloropsis sp. ,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

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