RbcL | Rubisco large subunit, form I and form II (100 µl)

352 €

AS03 037 | 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

PRODUCT INFORMATION IN PDF

product information

background

This antibody is especially suitable for quantifying of Rubisco in plant and algal samples.

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

antibody format

rabbit

polyclonal,

serum,

lyophilized

quantity

100 µl

for reconstitution add 100 µl of sterile water

storage

store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

tested applications

western blot (WB), tissue printing (TP), immunofluorescence/confocal microscopy (IF), immunolabelling (IL)

related products

AS03 037-10 anti-RbcL antibody, smaller pack size (10 µl) of AS03 037

AS03 037-200 anti-RbcL antibody, larger pack size (200 µl) of AS03 037

AS09 409 | Rubisco quantitation kit

AS01 017 anti-RbcL | Rubisco large subunit, form I

AS07 218 anti-Rubisco | 557 kDa hexadecamer

AS01 017S anti-RbcL | Rubisco protein standard

AS07 259 anti-RbcS | Rubisco small subunit (SSU)

AS07 222 RbcS | Rubisco small subunit (SSU) from pea

recommended secondary antibody

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.

Example of a simulataneous western blot detection with RbcL, PsbA and PsaC antibodies.

application information
recommended dilution		1: 5000 - 10 000 with standard ECL (WB), 1: 800 (TP), immunofluorescence/confocal microscopy (IF), 1: 250 for images see Prins et al. (2008), detailed protocol available on request (IL)
expected \| apparent MW		52.7 kDa (Arabidopsis thaliana), 52.5 kDa (cyanobacteria), 52.3 (Chlamydomonas reinhardtii)
confirmed reactivity		Arabidopsis thaliana, Apium graveolens, Baculogypsina sphaerulata (benthic foraminifer), Bienertia sinuspersici, Cicer arietinum, 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, Liquidambar formosana, Micromonas pusila, Nicotiana benthamiana, Physcomitrella patens, Porphyra sp. , Schima superba, Stanleya pinnata, Spinacia oleracea, lichens, Synechococcus PCC 7942, Thalassiosira pseudonana, 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
not reactive in		no confirmed exceptions from predicted reactivity known in the moment
additional information		This antibody was used in: Immunocytochemical staining of diatoms according to Schmid (2003) J Phycol 39: 139-153 and Wordemann et al. (1986) J Cell Biol 102: 1688-1698. Immunofluorescence Dreier et al. (2012). FEMS Microbial Ecol., March 2012. Western blot and tissue printing during a student course Ma et al. (2009).
selected references		Losh et al. (2013). Rubisco is a small fraction of total protein in marine phytoplankton. New Phytol. April 198 (1):52-8. Chen et al. (2013). Photosynthetic and antioxidant responses of Liquidambar formosana and Schima superba seedlings to sulfuric-rich and nitric-rich simulated acid rain. Plant Physiol & Biochem. Li at al. (2012). MAP Kinase 6-mediated activation of vacuolar processing enzyme modulates heat shock-induced programmed cell death in Arabidopsis. New Phytol. ahead of print - RbcL antibody used as loading control. Zhao et al. (2011). Expansins are involved in cell growth mediated by abscisic acid and indole-3-acetic acid under drought stress in wheat. Plant Cell Rep. Nov (RbcL antibody used as a loading control) Heckwolf et al. (2011). The Arabidopsis thaliana aquaporin AtPIP1;2 is a physiologically relevant CO(2) transport facilitator. Plant J. doi: 10.1111/j.1365-313X.2011.04634.x. [Epub ahead of print] Johnson (2011). Manipulating RuBisCO accumulation in the green alga, Chlamydomonas reinhardtii. Plant Mol Biol. May 24. Kubien et al. (2011). Quantification of the amount and activity of Rubisco in leaves. Methods Mol Biol. 2011;684:349-62. Nicolardi et al. (2011). The adaptive response of lichens to mercury exposure involves changes in photosynthetic machinery. Environmental Pollution (16): 1-10. Zilliges et al (2011) The Cyanobacterial Hepatotoxin Microcystin Binds to Proteins and Increases the Fitness of Microcystis under Oxidative Stress Conditions. PLoS ONE.

application example

0.25 µg of chlorophyl a/lane from Spinacia oleracea (1), Synechococcus PCC 7942 (2), Cyanophora paradoxa (3), Heterosigma akashiwo (4), Thalassiosira pseudonana (5), Euglena gracilis (6), Micromonas pusilla (7), Chlamydomonas reinhardtii (8), Porphyra sp (9), Gonyaulax polyedra (10), Emiliania huxleyi (11) extracted with PEB (AS08 300), were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to nitrocellulose. Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-RbcL antibody (AS03 037, 1:50 000, 1h) and secondary anti-rabbit (1:20000, 1 h) antibody (HRP conjugated, recommended secondary antibody AS09 602) in TBS-T containing 2% low fat milk powder. Antibody incubations were followed by washings in TBS-T. All steps were performed at RT with agitation. Blots were developed for 5 min with ECL Advance detection reagent according the manufacturers instructions (GE Healthcare). Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).

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