Recombinant Anti-p40-phox antibody [EP2142Y] (ab76158)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2142Y] to p40-phox
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-p40-phox antibody [EP2142Y]
See all p40-phox primary antibodies -
Description
Rabbit monoclonal [EP2142Y] to p40-phox -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human p40-phox aa 300-400 (C terminal). The exact sequence is proprietary.
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Positive control
- WB: HL60 cell lysate. IHC: human spleen tissue. IF/ICC: MCF7 cell line. IP: HL-60. Flow Cyt: K562 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2142Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab76158 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/20 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/100 - 1/250.
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WB | (1) |
1/25000 - 1/50000. Detects a band of approximately 39 kDa (predicted molecular weight: 39 kDa).
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IP |
1/50.
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IHC-P |
1/100 - 1/250.
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Notes |
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Flow Cyt (Intra)
1/20 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/100 - 1/250. |
WB
1/25000 - 1/50000. Detects a band of approximately 39 kDa (predicted molecular weight: 39 kDa). |
IP
1/50. |
IHC-P
1/100 - 1/250. |
Target
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Function
Component of the NADPH-oxidase, a multicomponent enzyme system responsible for the oxidative burst in which electrons are transported from NADPH to molecular oxygen, generating reactive oxidant intermediates. It may be important for the assembly and/or activation of the NADPH-oxidase complex. -
Tissue specificity
Expression is restricted to hematopoietic cells. -
Sequence similarities
Contains 1 PX (phox homology) domain.
Contains 1 SH3 domain. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 4689 Human
- Omim: 601488 Human
- SwissProt: Q15080 Human
- Unigene: 474781 Human
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Alternative names
- CGD3 antibody
- MGC3810 antibody
- NCF 4 antibody
see all
Images
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Anti-p40-phox antibody [EP2142Y] (ab76158) at 1/50000 dilution + HL60 cell lysate at 10 µg
Secondary
Goat anti-rabbit HRP conjugated at 1/2000 dilution
Predicted band size: 39 kDa
Observed band size: 39 kDa -
ICC/IF image of ab76158 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76158, 1/25 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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ab76158 at 1/100-1/250 dilution staining p40-phox in human spleen by Immunohistochemistry, Paraffin-embedded tissue.
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Overlay histogram showing K562 cells stained with ab76158 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76158, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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p40-phox was immunoprecipitated from 0.35 mg HL-60 (Human acute promyelocytic leukemia promyeloblast) whole cell lysate 10 µg with 76158 at 1/50 dilution (2µg). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HL-60 (Human acute promyelocytic leukemia promyeloblast) whole cell lysate 10 µg
Lane 2: ab76158 IP in HL-60 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab76158 in HL-60 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (5)
ab76158 has been referenced in 5 publications.
- Kuhns DB Diagnostic Testing for Chronic Granulomatous Disease. Methods Mol Biol 1982:543-571 (2019). PubMed: 31172495
- Merling RK et al. An AAVS1-targeted minigene platform for correction of iPSCs from all five types of chronic granulomatous disease. Mol Ther 23:147-57 (2015). PubMed: 25288370
- Lamb FS et al. Endotoxin priming of neutrophils requires endocytosis and NADPH oxidase-dependent endosomal reactive oxygen species. J Biol Chem 287:12395-404 (2012). WB, ICC/IF ; Human . PubMed: 22235113
- Ueyama T et al. Cooperation of p40phox with p47phox for Nox2-based NADPH Oxidase Activation during Fc? Receptor (Fc?R)-mediated Phagocytosis: MECHANISM FOR ACQUISITION OF p40phox PHOSPHATIDYLINOSITOL 3-PHOSPHATE (PI(3)P) BINDING. J Biol Chem 286:40693-705 (2011). PubMed: 21956105
- McCaffrey RL et al. Multiple mechanisms of NADPH oxidase inhibition by type A and type B Francisella tularensis. J Leukoc Biol 88:791-805 (2010). PubMed: 20610796