Anti-RbcL | Rubisco large subunit, form I, HRP-conjugated (40 µg)

Product no: AS03 037-HRP
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

Benefits of using this antibody
490
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Delivery:  3-6 business days
  • Product Info
  • 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
    Purity: Immunogen affinity purified serum in PBS pH 7.4, conjugated to HRP.
    Format: Liquid
    Quantity: 40 µg
    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)
    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)

  • Reactivity
  • Confirmed reactivity: Agostis stolonifera cv. Penncross, Arabidopsis thaliana, Apium graveolens, Artemisia annua, Atrichum undulatum, Attheya longicornis, Baculogypsina sphaerulata (benthic foraminifer), Beta vulgaris, Begonia sp., Bienertia sinuspersici, Brassica napus, Kandelia candel, Cannabis sativa L., Chaetoceros furcellatus, Chlorococcum dorsiventrale, Colobanthus quitensis, Cicer arietinum, Chenopodium quinoa, Chlamydomonas raudensis, Chlamydomonas reinhardtii, Colobanthus quitensis Kunt Bartl, Chlorella sorokiniana, Chlorella vulgaris, Coscinodiscus concinnus, Cyanophora paradoxa, Cylindrospermopsis raciborskii CS-505, Cynara cardunculus, Emiliana huxleyi, Euglena gracilis, Ficus carica, Fortunella margarita Swingle, Fraxinus mandshurica, Fucus vesiculosus, Gladieria sulphuraria, Glycine max, Gonyaulax polyedra, Guzmania hybrid, Heterosigma akashiwo, Hevea, Hordeum vulgare, Hypnum cupressiforme, Jatropha curcas, Karenia brevis (C.C.Davis) s) G.Hansen & Ř.Moestrup (Wilson isolate), Kochia prostrata, Lathyrus sativus, Liquidambar formosana, Malus domestica, Medicago truncatula, Micromonas pusila, Nicotiana benthamiana, Nicotiana tabacum, Panicum virgatum, Petunia hybrida cv. Mitchell, Phaeodactylum tricornutum, Physcomitrium patens, Pisum sativum, olytrichum formosum, Porosira glacialis,, Porphyra sp., Ricinus communis, Robinia pseudoacacia, Rhytidiadelphus squarrosus, Saccharum sp., Schima superba, Skeletonema costatum (diatom), Skeletonema marinoi (diatom), Solanum lycopersicum, Spinacia oleracea, lichens, Stanleya pinnata, Symbiodinium sp., Synechococcus PCC 7942, Synechococcus elongatus UTEX 2973, Rhoeo discolor, Thalassiosira pseudonana, Thermosynechococcus elongatus, Triticum aestivum, Prochlorococcus sp. (surface and deep water ecotype), Triticum aestivum, dinoflagellate endosymbionts (genus Symbiodinium), extreme acidophilic verrucomicrobial methanotroph Methylacidiphilum fumariolicum strain SolV, Thalassiosira punctigera, Tisochrysis lutea, Verbascum lychnitis, Vitis vinifera, Quercus ilex
    Predicted reactivity: Alpha proteobacteria, Algae (brown and red) including Galdieria sulphuraria, Dicots, Benincasa hispida, Kalanchoe fedtschenkoi; Beta-proteobacteria, Conifers, Cryptomonads, Cyanobacteria (prochlorophytes), Gamma-proeobacteria, Liverworts, Manihot esculenta, Marchantia polymorpha, Monocots, Mosses, Suaeda glauca, Welwitschia; Nannochloropsis sp., Picochlorum sp., Porphyridium purpureum, Zea mays, Zosteria marina

    For detection in Rhodospirillaceae use product AS15 2955
    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Additional Information
  • 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.
  • Background
  • 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.

    This antibody is directly conjugated to HRP.

  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts



    Rubisco quantitation in plant and algal samples using Agrisera anti-RbcL antibody and Rubisco protein standard

    Methodology: Plant samples are generally ground with liquid nitrogen in a mortar and pestle. The resulting powder is transferred to a plastic tube. Algal samples can be either concentrated by centrifugation or, preferably, by filtration onto glass fiber filters. Solubilization is performed in Agrisera protein extraction buffer (PEB, AS08 300) containing 0.1mg/mL PefaBloc SC (AEBSF) protease inhibitor (Roche). Disruption is most optimally obtained through flash freezing of the sample in liquid nitrogen alternated with thawing by sonication with a microtip. This process can be repeated depending on the toughness of the sample. The sample is adjusted to 50 mM dithiothreitol and heated to 70°C for 5 minutes. Samples are cooled and centrifuged briefly prior to electrophoresis.

    Optimal quantitation is achieved using moderate sample loads per gel lane, generally 0.5 to 2.5 ug total protein, depending on the abundance of the target protein.

    Electrophoresis and Immunoblotting: Once solubilized, the proteins can be separated electrophoretically in a number of systems. We obtain optimal results with the Invitrogen NuPAGE gel system using Bis-Tris 4-12% gradient gels. Proteins are separated in MES SDS running buffer according to the manufacturer’s recommendations at 200 V for 35 minutes. The gels are transferred to PVDF in the same apparatus, the SureLock XCell blot module, for 60 minutes at 30 V for a single gel or 80 minutes for a pair. Following transfer the blots are blocked with non-fat dry milk up to 10 % in TBS-T, for 1 h/RT with gentle agitation. The blot is incubated with primary antibody, usually at 1:25 000 to 1:50 000, for 1 h/RT (if extreme femtogram detection reagents are used) or in lower primary antibody dilution for less sensitivie reagents (mid picogram and lower).

    For quantitation a relatively high primary antibody: target protein ratio gives more reliable results than immunoblots at low ratios of primary antibody:target protein.

    The blot is washed extensively in TBS-T (twice briefly, once for 15 minutes and three times for five minutes). When directly HRP-conjugated primary antibody is used, there is no need for incubation with secondary antibody. The blot is washed as above and developed with ECL detection reagents.

    Quantitation: When quantitated standards are included on the blot, the samples can be quantitated using the available software. Excellent quantitation can be obtained with images captured on the Bio-Rad Fluor-S-Max or equivalent instrument using Bio-Rad QuantityOne software. The contour tool is used to select the area for quantitation and the values are background subtracted to give an adjusted volume in counts for each standard and sample. Using above protocol linear standard curves are generated over 1-1.5 orders of magnitude range in target load. It is important to note that immunodetections usually show a strongly sigmoidal signal to load response curve, with a region of trace detection of low loads, a pseudolinear range and a region of saturated response with high loads. For immunoquantitation it is critical that the target proteins in the samples and the standard curve fall within the pseudolinear range. Our total detection range using this protocol spans over 2 orders of magnitude, but the quantifiable range is narrower.


    References:

    MacKenzie et al  (2005). Large reallocations of carbon, nitrogen and photosynthetic reductant among phycobilisomes, photosystems and Rubisco during light acclimation in Synechococcus elongatus are constrained in cells under low environmental inorganic carbon. Arch of Microbiol. 183: 190 - 202.
    Bouchard et al. (2006) UVB effects on the photosystem II-D1 protein of phytoplankton and natural phytoplankton communities. Photochem and Photobiol 82: 936-951. 
    Morash
    et al. (2007) Macromolecular dynamics of the photosynthetic system over a seasonal developmental progression in Spartina alterniflora. Can J. of Bot. 85: 476-483(8)


    Recommended chemiluminescent detection reagent: AgriseraECL Bright (100 ml)

  • Reviews:
  • Reviews
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    Number of reviews: (1)
    Tomasz Hura | 2021-02-01
    0 | 0
    This antibody work very well in both triticale and Rosa rubiginosa

Accessories

AS03 037-ALP Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: [global antibody] for higher plants, lichens,  algae, cyanobacteria, dinoflagellates, diatoms compartment marker  of chloroplast stroma in higher plants and cytoplasm in cyanobacteria

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490 €
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AS03 037 | Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: global antibody  and compartment marker for higher plants, lichens, algae, cyanobacteria, dinoflagellates, diatoms 

Benefits of using this antibody

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