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RA | Rubisco activase

AS10 700  | Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: A. thaliana, C. sativa, C. reinhardtii, H. spontaneum, F. pRatensis, G. max, G. hirsutum, G. barbadense, L. perenne, N. oceanica, N. tabacum, O. sativa, P. balsamifera, R. discolor, S. lycopersicum, Z. mays, T. salsuginea, red sulfur bacterium T. sp. Cad16 (isolated from Lake Cadagno)

RA | Rubisco activase in the group Antibodies for Plant/Algal  / Photosynthesis  / RUBISCO/Carbon metabolism at Agrisera AB (Antibodies for research) (AS10 700)

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Data sheet Product citations Protocols Customer reviews

Product Information

Immunogen

Purified, recombinant Rubisco activase from Gossypium hirsutum Q9AXG1

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 µl
Reconstitution For reconstitution add 50 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)
Recommended dilution 1 : 5000-1 : 10 000 (WB)
Expected | apparent MW

47 and 42 kDa (maize, tobacco, Chlamydomonas)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Camelina sativa, Chlamydomonas reinhardtii, Hordeum spontaneum, Festuca pratensis, Glycine max, Gossypium hirsutum, Gossypium barbadense, Lolium perenne, Nannochloropsis oceanica, Nicotiana tabacum, Oryza sativa, Populus balsamifera, Rhoeo discolor, Solanum lycopersicum, Zea mays, Thellungiella salsuginea, red sulfur bacterium Thiodictyon sp. Cad16 (isolated from Lake Cadagno)
Predicted reactivity Glycine max, Gossypium mexicanum, Hordeum vulgare, Medicago sativa, Olea europea, Picea sitcHensis, Physcomitrella patens, Ricinus communis, Solanum lycopersicum, Spinacia oleracea, Triticum aestivum, Zea mays, Vitis vinifera
Species of your interest not listed? Contact us
Not reactive in marine picocyanobacteria

Application examples

Application examples

Application example
western blot using Rubisco activase antibody

3, 5 and 11 μg of total soluble protein from Arabidopsis thaliana (1), Oryza sativa (2) and Camelina sativa (3) extracted with 50 mM Tricine-NaOH, pH 8, 10 mM EDTA, 1% PVP-40, 20 mM β-mercaptoethanol, 1 mM PMSF and 10 μM leupeptin were separated on 12 % SDS-PAGE and blotted 1h to PVDF. Blots were blocked with 4% non-fat milk in TBS for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10 000 for over night with agitation. The antibody solution was decanted and the blot was rinsed briefly with H2O, then washed six times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG alkaline phosphatase conjugated) diluted to 1:3000 in 0.5% non-fat milk in TBS for 2h at RT with agitation. The blot was washed with four changes of TBS-T and developed for 5 min with NBT/BCIP according to the manufacturer’s instructions (Promega). There are two forms of activase (alpha and beta). Alpha is about 46-47 Kda, beta is about 42 kDa what is shown on the blot above.

western blot detection of Rubisco activase in various species
5 µg of total protein from samples such as Arabidopsis thaliana leaf (1) ,  Hordeum vulgare leaf (2)Zea mays leaf (3)Nicotiana tabacum (4), Chlamydomonas reinhardtii total cell (5), 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 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: 10 000 (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. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS10 1489, Agrisera) diluted to 1:25 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with chemiluminescent 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.

Additional information

Additional information This product can be sold containing ProClin if requested
There are two forms of activase (alpha and beta) in some species (for example Arabidopsis, camelina, spinach, rice) and only one form in other species (tobacco, maize, Chlamydomonas). Alpha is about 46-47 Kda, beta is about 42 kDa. Species that have only one form have the beta form.

This product can be sold containing ProClin if requested

Related products

Background

Background

RA - Ribulose bisphosphate carboxylase/oxygenase activase is an enzyme localized to chloroplasts which activates Rubisco by promoting ATP-dependent conformational changes. Alternative name: RuBisCo activase RCA.

Product citations

Selected references Suganami et al. (2020). Effects of Overproduction of Rubisco Activase on Rubisco Content in Transgenic Rice Grown at Different N Levels. Int J Mol Sci. 2020 Feb 27;21(5). pii: E1626. doi: 10.3390/ijms21051626.
Salesse-Smith et al. (2018). Overexpression of Rubisco subunits with RAF1 increases Rubisco content in maize. Nat Plants. 2018 Oct;4(10):802-810. doi: 10.1038/s41477-018-0252-4.
Yoshida et al. (2018). Thioredoxin-like2/2-Cys peroxiredoxin redox cascade supports oxidative thiol modulation in chloroplasts. Proc Natl Acad Sci U S A. 2018 Aug 13. pii: 201808284. doi: 10.1073/pnas.1808284115.
Tamburino et al. (2017). Chloroplast proteome response to drought stress and recovery in tomato (Solanum lycopersicum L.). BMC Plant Biol. 2017 Feb 10;17(1):40. doi: 10.1186/s12870-017-0971-0.
Wei et al. (2017). Enhancing photosynthetic biomass productivity of industrial oleaginous microalgae by overexpression of RuBisCO activase. Algal Research Volume 27, November 2017, Pages 366-375.
Yin et al. (2016). Interplay between mitogen-activated protein kinase and nitric oxide in brassinosteroid-induced pesticide metabolism in Solanum lycopersicum. J Hazard Mater. 2016 Oct 5;316:221-31. doi: 10.1016/j.jhazmat.2016.04.070. Epub 2016 Apr 29.
Hu et al. (2015). Site-specific Nitrosoproteomic Identification of Endogenously S-Nitrosylated Proteins in Arabidopsis. Plant Physiol. 2015 Feb 19. pii: pp.00026.2015.
Jurczyk et al. (2015). Evidence for alternative splicing mechanisms in meadow fescue (Festuca pratensis) and perennial ryegrass (Lolium perenne) Rubisco activase gene. J Plant Physiol. 2014 Dec 18;176C:61-64. doi: 10.1016/j.jplph.2014.11.011.
Jedmowski et al. (2014). Comparative analysis of drought stress effects on photosynthesis of Eurasian and North African genotypes of wild barley. Photosynthetica, September 2014.
Yin et al. (2014). Characterization of Rubisco activase genes in maize: an α-isoform gene functions alongside a β-isoform gene. Plant Physiol. 2014 Apr;164(4):2096-106. doi: 10.1104/pp.113.230854. Epub 2014 Feb 7.
Wiciarz et al. (2014). Enhanced chloroplastic generation of H2 O2 in stress-resistant Thellungiella salsuginea in comparison to Arabidopsis thaliana. Physiol Plant. 2014 Jun 24. doi: 10.1111/ppl.12248.
Chen et al. (2013). Physiological Mechanisms for High Salt Tolerance in Wild Soybean (Glycine soja) from Yellow River Delta, China: Photosynthesis, Osmotic Regulation, Ion Flux and antioxidant Capacity. PLoS One. 2013 Dec 12;8(12):e83227. doi: 10.1371/journal.pone.0083227.

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