PsbA | D1 protein of PSII, C-terminal (affinity purified antibody)
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1: 15 000 with standard ECL (WB)
|Expected | apparent MW||
38 | 28-30 kDa
Arabidopsis thaliana, Artemisia annua, Arundo sp., Colobanthus quitensis Kunt Bartl, Craterostigma sp. Glycine max, Hordeum vulgare, Lindernia sp., Miscanthus x giganteus, Marchantia polymorpha (liverwort), Nicotiana benthamiana,Panicum miliaceum, Panax ginseng, Panicum maximum, Pinus strobus, Zea mays, Physcomitrella patens, Chlamydomonas reinhardtii, Synechococcus sp. PCC 7942, Anabaena 7120, Paulinella chromatophora (amoeba), Prochlorococcus sp. (surface and deep water ecotype), Spirodela polyrhiza, Symbiodinium sp., Coscinodiscus wailesii, Ditylum brightwellii
Medicago sativa, Pisum sativum, Triticum aestivum and other di and monocots,conifers, brown and red algae, cyanobacteria; cellular [compartment marker] of thylakoid membrane
|Not reactive in||
no confirmed exceptions from predicted reactivity known in the moment
The antibody is appropriate for detecting both, 24 kDa or the 10 kDa C-terminal fragments, whichever is generated under given treatment conditions. In our analysis we have seen both, ca. 24 kDa and ca. 10 kDa fragments from different samples, depending on treatments and isolation procedures.
Rabbit anti-PsbA antibody can detect more than one band of PsbA protein, e.g. precursor and mature protein as compare to the hen anti-PsbA antibodies AS01 016.
This antibody will detect the phosphorylated form of D1 as an alternate band to the main band on a high resolution gel.
|Selected references||to be added when available, antibody released in March 2015|
2 µg of total protein from samples such as Arabidopsis thaliana leaf (1) , Hordeum vulgare leaf (2), Chlamydomonas reinhardtii total cell (3), Synechococcus sp. PCC7942 total cell (4), 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 keept on ice before loading. Protein samples were separated on 4-12% Bolt Plus gels, LDS-PAGE and blotted for 70 minutes to PVDF using tank transfer. Blots were blocked immediately following transfer in 2% blocking reagent (GE RPN 2125; Healthcare) or 5% non-fat milk dissolved 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 (in blocking reagent) for 1h at room temperature 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 room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS09 602, Agrisera) diluted to 1:25 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with ECL detection reagent according to the manufacturer's instructions. Images of the blots were obtained using a CCD imager (VersaDoc MP 4000) and Quantity One software (Bio-Rad). Exposure time was 30 seconds.
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