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Plant/Algal cell antibodies / Conifers


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

Art no: AS03 037
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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. 

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, Bienertia sinuspersici, Chlamydomonas reinhardtii, Cyanophora paradoxa, Emiliana huxleyi, Euglena gracilis, Fraxinus mandshurica, Glycine max, Gonyaulax polyedra, Guzmania hybrid, Heterosigma akashiwo, Micromonas pusila, Nicotiana benthamiana, Physcomitrella patens, Porphyra sp. , 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

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  

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

RbcL western blot

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). 


||| For applications or usage on species others than stated above Agrisera offers a payment-after-testing option. To learn more about this or for any questions on this product, please use the LiveChat option in the left menue bar or contact us at  support@agrisera.com



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