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



DATA SHEET IN PDF

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Product Information

Immunogen

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

Host Rabbit
Clonality Polyclonal
Purity Immunogen 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 the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
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, Cucumis melo, Fragaria ananassa, Gossypium arboreum, Manihot esculenta, Morus notabilis, Nicotiana tabacum, Oryza sativa, Picea abies, Ricinus communis, Sinensis osbeck, Solanum tuberosum, Theobroma cacao, Zea mays, Zostera marina
Species of your interest not listed? Contact us
Not reactive in Marchantia polymorpha

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

Related products

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 Wang et al. (2021) Arabidopsis PUB2 and PUB4 connect signaling components of pattern-triggered immunity. New Phytol. 2021 Dec 17. doi: 10.1111/nph.17922. Epub ahead of print. PMID: 34918346
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.
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.
Otulak-Kozie? 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)
immunogen:

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

Reconstitution: For reconstitution add 50 µl of sterile water
Host: Rabbit
Clonality: Polyclonal
Purity: Immunogen affinity purified serum in PBS pH 7.4.
Format: Lyophilized
Quantity: 50 µg
storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications: Immunofluorescence (IF), Western blot (WB)
recommended dilution: 1 : 1000 (WB)
calculated | apparent molecular mass [kDa]:

104 kDa

Confirmed reactivity: Arabidopsis thaliana, Solanum sp.
predicted reactivity: Brassica napus, Citrus sinensis, Citrullus colocynthis, Cucumis melo, Fragaria ananassa, Gossypium arboreum, Manihot esculenta, Morus notabilis, Nicotiana tabacum, Oryza sativa, Picea abies, Ricinus communis, Sinensis osbeck, Solanum tuberosum, Theobroma cacao, Zea mays, Zostera marina
Species of your interest not listed? Contact us
not reactive in: Marchantia polymorpha
Picture (footer):

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 (application): For solubilization we recommend 65°C for 5 min, Sample boiling is not recommended
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.
All references: Wang et al. (2021) Arabidopsis PUB2 and PUB4 connect signaling components of pattern-triggered immunity. New Phytol. 2021 Dec 17. doi: 10.1111/nph.17922. Epub ahead of print. PMID: 34918346
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.
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.
Otulak-Kozie? 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)

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