RA | Rubisco activase

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AS10 700  | clonality: polyclonal  |  host: rabbit  |  reactivity: A. thaliana, C. sativa, C. reinhardtii, F. pratensis, G. max, G. hirsutum, L. perenne, N. tabacum, O. sativa, P. balsamifera, R. discolor, Thiodictyon sp., T. salsuginea, Z.mays


31 st
Item No:
AS10 700

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product information

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.


Purified, recombinant Rubisco activase from Gossypium hirsutum Q9AXG1

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 ĩl
Reconstitution For reconstitution add 50 ĩl of sterile water.

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)
Related products

Antibodies to RbcL, RbcS

Plant and algal protein extraction buffer

Secondary antibodies

Additional information
application information
Recommended dilution

1 : 5000 - 1: 10 000 with NBT/BCIP (WB)

Expected | apparent MW

47 and 42 kDa (maize, tobacco, Chlamydomonas)

Confirmed reactivity

Arabidopsis thaliana, Camelina sativa, Chlamydomonas reinhardtii, Hordeum spontaneum, Festuca pratensis, Glycine max, Gossypium hirsutum, Gossypium barbadense, Lolium perenne, Nicotiana tabacum, Oryza sativa, Populus balsamifera, Rhoeo discolor, Zea mays, Thellungiella salsuginea, red sulfur bacterium Thiodictyon sp. Cad16 (isolated from Lake Cadagno)

Predicted reactivity

Dicots including: Gossypium mexicanum, Glycine max, Medicago sativa, Ricinus communis, Spinacia oleracea, Olea europea, Picea sitchensis, Vitis vinifera; monocots. Hordeum vulgare, Triticum aestivum, Zea mays, moss: Physcomitrella patens

Not reactive in

no confirmed exceptions from predicted reactivity known in the moment

Additional information 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.
Selected references 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.
 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.

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, from Sigma-Aldrich) 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 (GE RPN 2125; Healthcare) 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 TMA-6 (Lumigen) 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.

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