| | product information | | Background | | Rubisco (Ribulose-1,5-bisphosphate carboxylase/oxygenase) catalyzes the rate-limiting step of CO2 fixation in photosynthetic organisms. It is demonstrably homologous from purple bacteria to flowering plants and consists of two protein subunits, each present in 8 copies. In plants and green algae, the large subunit (~55 kDa) is coded by the chloroplast rbcL gene, and the small subunit (15 kDa) is coded by a family of nuclear rbcS genes. | | Immunogen | | KLH-conjugated synthetic peptide conserved across all known plant, algal and (cyano)bacterial RbcL protein sequences (form I L8S8 and form II L2), including Arabidopsis thaliana AtCg00490, Hordeum vulgare P05698, Oryza sativa P0C510, Chlamydomonas reinhardtii P00877, Synechococcus PCC 7920 A5CKC5 | | Host | | Rabbit | | Clonality | | Polyclonal | | Clone | | | | Purity | | Affinity purified serum | | Format | | Liquid in PBS pH 7.4, conjugated to HRP | | Quantity | | 40 µg | | Reconstitution | | | | Storage | | Store at +4°C for 12-18 months. A preservative may be added for long time storage up to 2 years. | | Tested applications | | ELISA (ELISA), western blot (WB) | | Related products | | AS03 037 | anti-RbcL | Rubisco large subunit, form I and form II (50 µl)AS03 037A | anti-RbcL | Rubisco large subunit, form I and form II (50 µg affinity purified)AS15 2955 | anti-RbcL II | Rubisco large subunit, form II (50 µl), rabbit antibodyAS15 2955S | RbcL II | Rubisco form II positive control/quantitation standardAS01 017 | anti-RbcL | Rubisco large subunit, form I, chicken antibodyAS01 017S | Rubisco protein standard for quantitative western blot or positive controlAS03 037PRE | Rubisco large subunit, pre-immune serumAS09 409 | Rubisco quantitation kit AS15 2994 | Rubisco ELISA quantitation kit AS07 218 | anti-Rubisco | 557 kDa hexadecamer, rabbit antibody to a whole protein AS07 259 | anti-RbcS | Rubisco small subunit (SSU), rabbit antibodyAS07 222 | anti-RbcS | Rubisco small subunit (SSU) from pea, rabbit antibody matching secondary antibody Plant and algal protein extraction buffer Secondary antibodies | | Additional information | | anti-RbcL can be used as a cellular [compartment marker] of plastid stroma (cytoplasm in cyanobacteria) and detects RbcL protein from 31.25 fmoles. As both forms (I and II) are detected it is suitable for work with samples from Dinoflagellates, Haptophytes and Ochrophytes (diatoms, Raphidophytes, brown algae) as well as higher plants. This antibody together with Agrisera Rubisco protein standard is very suitable to quantify Rubisco in plant and algal samples. | | application information | | 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 | | di and monocots, conifers, mosses, liverworts, welwitschia, green algae, red alge, brown algae, cryptomonad, cyanobacteria including prochlorophytes, gamma-proeobacteria, beta-proteobacteria, alpha proteobacteria, Suaeda glaucaPredicted but not confirmed reactivity for Rubisco form II. | | Not reactive in | | no confirmed exceptions from predicted reactivity known in the moment | | Additional information | | | | Selected references | | | Application example![\"western](\"https://www.ebiomall.com/images/agrisera/201709/AS03) 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.
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