PsaD | PSI-D subunit of photosystem I
AS09 461 | clonality: polyclonal | host: rabbit | reactivity: plants (monocots and dicots, conifers), moss: Physcomitrella patens, Chlamydomonas reinhardtii
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1 : 1000 (WB)
|Expected | apparent MW||
17.9 | 20 (for Arabidopsis thaliana)
Arabidopsis thaliana, Chlamydomonas reinhardtii, Hordeum vulgare, Lactuca sativa, Oryza sativa, Physcomitrella patens, Spinacia oleracea, Synechocystis PCC 6803, Triticum aestivum, Zea mays
plants (monocots, dicots and conifers), Bigelowiella natans, green algae
|Not reactive in||
Synechococcus elongatus sp. PCC 7942
this antibody is a replacement for former product, anti-PsaD AS04 046
|Selected references||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.
Gerotto et al. (2016). Flavodiiron proteins act as safety valve for electrons in Physcomitrella patens. PNAS DOI 10.1073.
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
Fujii et al. (2015). Photoprotection vs Photoinhibition of Photosystem II in Transplastomic Lettuce (Lactuca sativa) Dominantly Accumulating Astaxanthin. Plant Cell Physiol. 2015 Dec 7. pii: pcv187.
Daddy et al. (2015). A novel high light-inducible carotenoid-binding protein complex in the thylakoid membranes of Synechocystis PCC 6803. Sci Rep. 2015 Mar 30;5:9480. doi: 10.1038/srep09480.
Qin et al. (2014). Isolation and characterization of a PSI-LHCI super-complex and its sub-complexes from a siphonaceous marine green alga, Bryopsis Corticulans. Photosynth Res. 2014 Sep 12.
Cheng and He (2014). PfsR Is a Key Regulator of Iron Homeostasis in Synechocystis PCC 6803. PLoS One. 2014 Jul 10;9(7):e101743. doi: 10.1371/journal.pone.0101743. eCollection 2014.
Tomizioli et al. (2014). Deciphering thylakoid sub-compartments using a mass spectrometry-based approach. Mol Cell Proteomics. 2014 May 28. pii: mcp.M114.040923.
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, Abcam) 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 SuperSignal West Pico (Thermo Scientific) 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 65oC 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, AS09 602) diluted to 1:20 000 for 1h at RT with agitation. The blots were washed as above, developed for 5 min with SuperSignal West Pico Chemiluminescent Substrate (34080, Thermo Scientific) 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
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