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RBOHD | Respiratory burst oxidase homolog protein D

AS15 2962  | Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: Arabidopsis thaliana, Solanum lycopersicum

RBOHD | Respiratory burst oxidase homolog protein D  in the group Antibodies for Plant/Algal  / Environmental Stress / Oxidative stress at Agrisera AB (Antibodies for research) (AS15 2962)

DATA SHEET IN PDF

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How to cite this product:
Product name, number (Agrisera, Sweden)

Data sheet Product citations Protocols Customer reviews

Product Information

Immunogen

KLH-conjugated peptide chosen from Arabidopsis thaliana RBOHD sequence, UniProt:Q9FIJ0, TAIR: AT5G47910

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized
Quantity 50 µg
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 Immunofluorescence (IF), Western blot (WB)
Recommended dilution 1 : 1000 (WB)
Expected | apparent MW

104 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Solanum sp.
Predicted reactivity Brassica napus, Citrus sinensis, Citrullus colocynthis, Fragaria ananassa, Gossypium arboreum, Manihot esculenta, Morus notabilis, Nicotiana tabacum, Oryza sativa, Picea abies, Ricinus communis, Solanum tuberosum, Theobroma cacao, Zea mays, Zostera marina
Species of your interest not listed? Contact us

Application examples

Application examples

Application example


Western blot using anti-RBOHD antibodies



Total proteins were isolated from 10 or 12 seedlings of 7-day-old Arabidopsis thaliana seedlings of Col-0 (wild-type) and rbohd null mutant (Nühse et al., 2007). 16 µl of total proteins were denatured at 65°C for 5 min, separated on an 8 % SDS-PAGE. and transferred for 70 min at 55V using a tank transfer system to nitrocellulose membrane. Blots were blocked with 1xPBS+0.1% Tween 20 + 5 % milk (PBS-T) or TBS + 0.1 % Tween 20 (TBS-T) + 5 % milk for 2 h at room temperature (RT) with agitation. Primary antibodies were diluted as indicated to 1: 2,500 and 1: 5,000 respectively and incubated ON at 4ºC with agitation in 1xPBS-T pr TBS-T + 5 % milk. The primary antibody solutions were decanted, and the blots were washed 3 times (7 min. each) in 1xPBS-T or TBS-T at RT with agitation. Blot was incubated with secondary antibody goat anti-rabbit HRP conjugated (AS09 602, Agrisera) diluted to 1: 20, 000 and developed with chemiluminescent detection reagent, accodring to manufacture's recommendations. Exposure time was 2 min. for X-ray film. As a control part of the same membrane was incubated with anti-CNX1/2 antibodies (AS12 2365, Agrisera).

References: 
Nühse et al. Quantitative phosphoproteomic analysis of plasma membrane proteins reveals regulatory mechanisms of plant innate immune responses. Plant J. 2007 Sep; 51(5): 931–940.

Courtesy of Nga Nguyen, Kelly Mason and Antje Heese (University of Missouri- Columbia, MO, USA

Additional information

For solubilization we recommend 65°C for 5 min. Sample boiling is not recommended. 

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Background

Background RBOHD (Respiratory burst oxidase homolog protein D) is a Calcium-dependent NADPH oxidase that generates superoxide. Is involved in the generation of reactive oxygen species (ROS) during incompatible interactions with pathogens and in UV-B and abscisic acid ROS-dependent signaling.

Product citations

Selected references Lee et al. (2020). Regulation of reactive oxygen species during plant immunity through phosphorylation and ubiquitination of RBOHD. Nat Commun. 2020 Apr 15;11(1):1838. doi: 10.1038/s41467-020-15601-5.
Otulak-Kozieł et al. (2019). The Respiratory Burst Oxidase Homolog D (RbohD)Cell and Tissue Distribution in Potato–Potato Virus Y (PVYNTN) Hypersensitive and Susceptible Reactions. Int. J. Mol. Sci. 2019, 20(11), 2741. (innunofluorescence)
Jedelská et al. (2019). Tomato Root Growth Inhibition by Salinity and Cadmium Is Mediated By S-Nitrosative Modifications of ROS Metabolic Enzymes Controlled by S-Nitrosoglutathione Reductase. Biomolecules. 2019 Aug 21;9(9). pii: E393. doi: 10.3390/biom9090393.

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