RA | Rubisco activase
AS10 700 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, Caesalpinia pulcherrima, 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)
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Purified, recombinant Rubisco activase from Gossypium hirsutum Q9AXG1
47 and 42 kDa (maize, tobacco, Chlamydomonas)
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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.
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.
Reactant: Nannochloropsis gaditana
Application: Western Blotting
Pudmed ID: 31297156
Journal: Biotechnol Biofuels
Figure Number: 6B
Published Date: 2019-07-13
First Author: Wei, L., El Hajjami, M., et al.
Impact Factor: 5.839Open Publication
Enzymatic assays of key biochemical CCM genes in N. oceanica under VLC/HC. a Enzyme activities of C4-like genes under VLC and HC, including ME, MDH, PEPC and PEPCK. b Quantification of PEPCK protein under VLC and HC. Immunoblot band was detected from the cellular extract under VLC but not under HC (loading of 15 ?g total protein)
This product can be sold containing ProClin if requested
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.
Oikawa et al. (2021) Mitochondrial movement during its association with chloroplasts in Arabidopsis thaliana. Commun Biol. 2021 Mar 5;4(1):292. doi: 10.1038/s42003-021-01833-8. PMID: 33674706.
Wang et al. (2021) Insights Into the Gene Regulation in Jasmonate-Induced Whole-Plant Senescence of Tobacco Under Non-Starvation Condition. Plant Cell Physiol. 2021 Sep 15:pcab140. doi: 10.1093/pcp/pcab140. Epub ahead of print. PMID: 34523687.
Trojak et al. (2021) Effects of partial replacement of red by green light in the growth spectrum on photomorphogenesis and photosynthesis in tomato plants. Photosynth Res. 2021 Sep 27. doi: 10.1007/s11120-021-00879-3. Epub ahead of print. PMID: 34580802.
Yokochi et al.(2021) Oxidative regulation of chloroplast enzymes by thioredoxin and thioredoxin-like proteins in Arabidopsis thaliana. Proc Natl Acad Sci U S A. 2021 Dec 21;118(51):e2114952118. doi: 10.1073/pnas.2114952118. PMID: 34907017; PMCID: PMC8713810.
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.
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.
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.
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 ?-isoform gene functions alongside a ?-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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