RbcL | Rubisco large subunit, form I and form II (40 ĩg, HRP-conjugated)
AS03 037-HRP | clonality: polyclonal | host: rabbit | reactivity: [global antibody] for higher plants, lichens, algae, cyanobacteria, dinoflagellates, diatoms | cellular [compartment marker] of plastid stroma in higher plants and cytoplasm in cyanobacteria
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|Recommended dilution||1 : 5000 (ELISA), 1 : 10 000-25 000 (WB)|
|Expected | apparent MW||
52.7 kDa (Arabidopsis thaliana), 52.5 kDa (cyanobacteria), 52.3 (Chlamydomonas reinhardtii)
|Confirmed reactivity||Arabidopsis thaliana, Apium graveolens, Artemisia annua, Baculogypsina sphaerulata (benthic foraminifer), Bienertia sinuspersici, Cicer arietinum, Chlamydomonas raudensis, Chlamydomonas reinhardtii, Colobanthus quitensis Kunt Bartl, Cyanophora paradoxa, Cylindrospermopsis raciborskii CS-505, Emiliana huxleyi, Euglena gracilis, Fraxinus mandshurica, Fucus vesiculosus, Glycine max, Gonyaulax polyedra, Guzmania hybrid, Heterosigma akashiwo, Karenia brevis (C.C.Davis) s) G.Hansen & Ø.Moestrup (Wilson isolate), Liquidambar formosana, Micromonas pusila, Nicotiana benthamiana, Physcomitrella patens, Porphyra sp. , Schima superba, Stanleya pinnata, Spinacia oleracea, lichens, Symbiodinium sp., Synechococcus PCC 7942, Thalassiosira pseudonana, Thermosynechococcus elongatus, Prochlorococcus sp. (surface and deep water ecotype), Triticum aestivum, dinoflagellate endosymbionts (genus Symbiodinium), extreme acidophilic verrucomicrobial methanotroph Methylacidiphilum fumariolicum strain SolV, Thalassiosira punctigera, Vitis vinifera|
|Predicted reactivity||Aalpha proteobacteria, Algae (brown and red), Dicots, Beta-proteobacteria, Conifers, Cryptomonads, Cyanobacteria (prochlorophytes), Gamma-proeobacteria, Liverworts, Monocots, Mosses, Suaeda glauca, Welwitschia|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
1 µg of total protein from samples such as Arabidopsis thaliana leaf (1) , Hordeum vulgare leaf (2), Zea mays leaf (3), Chlamydomonas reinhardtii 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: 25 000 (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. The blot was developed for 5 min with TMA-6 (Lumigen) detection reagent according the manufacturers 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.
Rubisco protein from barley is visualized as two bands.