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AB76158

Anti-p40-phox antibody [EP2142Y]

5

(1 Review)

|

(8 Publications)

Rabbit Recombinant Monoclonal p40-phox antibody. Suitable for IP, WB, IHC-P, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 8 publications.

View Alternative Names

SH3PXD4, NCF4, Neutrophil cytosol factor 4, NCF-4, Neutrophil NADPH oxidase factor 4, SH3 and PX domain-containing protein 4, p40-phox, p40phox

5 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p40-phox antibody [EP2142Y] (AB76158)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p40-phox antibody [EP2142Y] (AB76158)

ab76158 at 1/100-1/250 dilution staining p40-phox in human spleen by Immunohistochemistry Paraffin-embedded tissue.

Immunocytochemistry/ Immunofluorescence - Anti-p40-phox antibody [EP2142Y] (AB76158)
  • ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-p40-phox antibody [EP2142Y] (AB76158)

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.

Flow Cytometry (Intracellular) - Anti-p40-phox antibody [EP2142Y] (AB76158)
  • Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-p40-phox antibody [EP2142Y] (AB76158)

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 >5000 events was performed.

Immunoprecipitation - Anti-p40-phox antibody [EP2142Y] (AB76158)
  • IP

Lab

Immunoprecipitation - Anti-p40-phox antibody [EP2142Y] (AB76158)

p40-phox was immunoprecipitated from 0.35 mg HL-60 (Human acute promyelocytic leukemia promyeloblast) whole cell lysate 10 μg with ab76158 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.

All lanes:

Immunoprecipitation - Anti-p40-phox antibody [EP2142Y] (ab76158)

Predicted band size: 39 kDa

Observed band size: 39 kDa

false

Western blot - Anti-p40-phox antibody [EP2142Y] (AB76158)
  • WB

Unknown

Western blot - Anti-p40-phox antibody [EP2142Y] (AB76158)

All lanes:

Western blot - Anti-p40-phox antibody [EP2142Y] (ab76158) at 1/50000 dilution

All lanes:

HL60 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP conjugated at 1/2000 dilution

Predicted band size: 39 kDa

Observed band size: 39 kDa

false

  • 578 PE

    PE Anti-p40-phox antibody [EP2142Y]

  • 660 APC

    APC Anti-p40-phox antibody [EP2142Y]

  • HRP

    HRP Anti-p40-phox antibody [EP2142Y]

  • 617 Alexa Fluor® 594

    Alexa Fluor® 594 Anti-p40-phox antibody [EP2142Y]

  • 519 Alexa Fluor® 488

    Alexa Fluor® 488 Anti-p40-phox antibody [EP2142Y]

  • 603 Alexa Fluor® 568

    Alexa Fluor® 568 Anti-p40-phox antibody [EP2142Y]

  • 565 Alexa Fluor® 555

    Alexa Fluor® 555 Anti-p40-phox antibody [EP2142Y]

  • Carrier free

    Anti-p40-phox antibody [EP2142Y] - BSA and Azide free

  • 665 Alexa Fluor® 647

    Alexa Fluor® 647 Anti-p40-phox antibody [EP2142Y]

  • 775 Alexa Fluor® 750

    Alexa Fluor® 750 Anti-p40-phox antibody [EP2142Y]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EP2142Y

Isotype

IgG

Carrier free

No

Reacts with

Human

Applications

Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/50", "IP-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/25000 - 1/50000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/100 - 1/250", "IHCP-species-notes": "<p></p>", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/100 - 1/250", "ICCIF-species-notes": "<p></p>", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/20 - 1/100", "FlowCytIntra-species-notes": "<p><a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a> - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.</p>" } } }
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Product details

Species reactivity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species.
Please contact us for more information.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 50% Tissue culture supernatant, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

P40-phox also known as neutrophil cytosolic factor 4 (NCF4) is a regulatory component of the NADPH oxidase complex. It is expressed in phagocytic cells such as neutrophils and macrophages. The protein plays a role in the production of reactive oxygen species (ROS) during the respiratory burst. The molecular mass of p40-phox is approximately 40 kDa. It interacts with other subunits of the NADPH oxidase complex such as p67-phox and p47-phox contributing to the assembly and activation of the complex.
Biological function summary

P40-phox participates in innate immune responses by regulating the NADPH oxidase complex's function. It acts as a scaffold to facilitate the interaction between cytosolic and membrane components. p40-phox integrates into the cytosolic fraction with other subunits and translocates to the membrane upon stimulation. This translocation is essential for generating superoxide radicals that are involved in defense against pathogens. Its functional domains enable it to bind phosphatidylinositol (34) bisphosphate which anchors the protein to the membrane. p40-phox itself is not an enzyme but modulates the activity of the complex.

Pathways

The p40 protein serves as an important player in the signaling cascade of the respiratory burst. It interacts with the phosphoinositide 3-kinase (PI3K) pathway and cooperates with other proteins like p67-phox and p47-phox through phosphorylation-dependent mechanisms. The p40-phox is phox compatible meaning it can integrate into various signaling modules. Within the oxidative burst pathway it enhances the cytosolic complex's attachment to the membrane affecting the production of ROS.

Mutations in the p40-phox have been connected to chronic granulomatous disease (CGD) characterized by recurrent infections and granuloma formation. The lack of functional p40-phox disrupts the assembly of the NADPH oxidase complex leading to decreased ROS production. This deficiency impairs the ability of phagocytes to eliminate pathogens effectively. p40-phox is also related to inflammatory disorders like inflammatory bowel disease (IBD) as variations in the NCF4 gene affect the immune response. The dysfunction in ROS production impacts the signaling involving p67-phox and p47-phox further influencing disease pathogenesis.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

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.
See full target information NCF4
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Publications (8)

Recent publications for all applications. Explore the full list and refine your search

Nature microbiology 9:2970-2984 PubMed39242815

2024

Mycobacterium tuberculosis virulence lipid PDIM inhibits autophagy in mice.

Applications

Unspecified application

Species

Unspecified reactive species

Ekansh Mittal,G V R Krishna Prasad,Sandeep Upadhyay,Jully Sadadiwala,Andrew J Olive,Guozhe Yang,Christopher M Sassetti,Jennifer A Philips

Nature communications 15:5170 PubMed38886341

2024

NCF4 attenuates colorectal cancer progression by modulating inflammasome activation and immune surveillance.

Applications

Unspecified application

Species

Unspecified reactive species

Longjun Li,Rudi Mao,Shenli Yuan,Qingqing Xie,Jinyu Meng,Yu Gu,Siyu Tan,Xiaoqing Xu,Chengjiang Gao,Hongbin Liu,Chunhong Ma,Si Ming Man,Xiangbo Meng,Tao Xu,Xiaopeng Qi

Annals of medicine and surgery (2012) 86:2644-2650 PubMed38694326

2024

S100A6 could not promote the differentiation of Calu-6 lung cancer cell line.

Applications

Unspecified application

Species

Unspecified reactive species

Jie Li,Ting Wang,Dan Dang

Methods in molecular biology (Clifton, N.J.) 1982:543-571 PubMed31172495

2019

Diagnostic Testing for Chronic Granulomatous Disease.

Applications

Unspecified application

Species

Unspecified reactive species

Douglas B Kuhns

Molecular therapy : the journal of the American Society of Gene Therapy 23:147-57 PubMed25288370

2014

An AAVS1-targeted minigene platform for correction of iPSCs from all five types of chronic granulomatous disease.

Applications

Unspecified application

Species

Unspecified reactive species

Randall K Merling,Colin L Sweeney,Jessica Chu,Aaron Bodansky,Uimook Choi,Debra Long Priel,Douglas B Kuhns,Hongmei Wang,Sam Vasilevsky,Suk See De Ravin,Thomas Winkler,Cynthia E Dunbar,Jizhong Zou,Kol A Zarember,John I Gallin,Steven M Holland,Harry L Malech

The Journal of biological chemistry 287:12395-404 PubMed22235113

2012

Endotoxin priming of neutrophils requires endocytosis and NADPH oxidase-dependent endosomal reactive oxygen species.

Applications

WB, ICC/IF

Species

Human, Human

Fred S Lamb,Jessica S Hook,Brieanna M Hilkin,Jody N Huber,A Paige Davis Volk,Jessica G Moreland

The Journal of biological chemistry 286:40693-705 PubMed21956105

2011

Cooperation of p40(phox) with p47(phox) for Nox2-based NADPH oxidase activation during Fcγ receptor (FcγR)-mediated phagocytosis: mechanism for acquisition of p40(phox) phosphatidylinositol 3-phosphate (PI(3)P) binding.

Applications

Unspecified application

Species

Unspecified reactive species

Takehiko Ueyama,Junya Nakakita,Takashi Nakamura,Takeshi Kobayashi,Toshihiro Kobayashi,Jeonghyun Son,Megumi Sakuma,Hirofumi Sakaguchi,Thomas L Leto,Naoaki Saito

Journal of leukocyte biology 88:791-805 PubMed20610796

2010

Multiple mechanisms of NADPH oxidase inhibition by type A and type B Francisella tularensis.

Applications

Unspecified application

Species

Unspecified reactive species

Ramona L McCaffrey,Justin T Schwartz,Stephen R Lindemann,Jessica G Moreland,Blake W Buchan,Bradley D Jones,Lee-Ann H Allen
View all publications
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Product promise

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