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Rabbit Recombinant Monoclonal NCF1/p47-phox antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with Mouse, Rat samples. Cited in 1 publication.

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Images

Flow Cytometry (Intracellular) - Anti-NCF1/p47-phox antibody [EPR27205-231] (AB308256), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-NCF1/p47-phox antibody [EPR27205-231] (AB308256), expandable thumbnail
  • Western blot - Anti-NCF1/p47-phox antibody [EPR27205-231] (AB308256), expandable thumbnail
  • Western blot - Anti-NCF1/p47-phox antibody [EPR27205-231] (AB308256), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IFFlow Cyt (Intra)IHC-PIP
Human
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Mouse
Tested
Tested
Tested
Not recommended
Not recommended
Rat
Tested
Expected
Expected
Not recommended
Not recommended

Tested
Tested

Species
Mouse
Dilution info
1/1000
Notes

-

Species
Rat
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/250
Notes

-

Expected
Expected

Species
Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/500
Notes

-

Expected
Expected

Species
Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat, Human
Dilution info
-
Notes

-

Target data

Function

NCF2, NCF1, and a membrane bound cytochrome b558 are required for activation of the latent NADPH oxidase (necessary for superoxide production).

Alternative names

Neutrophil cytosol factor 1, NCF-1, 47 kDa neutrophil oxidase factor, NCF-47K, Neutrophil NADPH oxidase factor 1, p47-phox, Ncf1

Recommended products

Rabbit Recombinant Monoclonal NCF1/p47-phox antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with Mouse, Rat samples. Cited in 1 publication.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR27205-231
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
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

Notes

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

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.

Supplementary info

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

NCF1 also known as p47-phox p47-phox protein or NCF-1 is a cytosolic component of the NADPH oxidase complex. It has a molecular mass of about 47 kDa. NCF1/p47-phox is expressed in neutrophils monocytes and other phagocytic cells. Functionally it plays an important role in the regulation of reactive oxygen species (ROS) production by facilitating the translocation of other subunits to the cell membrane where the NADPH oxidase complex assembles and becomes active.

Biological function summary

The NCF1/p47-phox protein plays a critical role in host defense by generating ROS which are essential for microbial killing. NCF1 is an integral part of the NADPH oxidase complex along with other subunits such as p67-phox p40-phox gp91-phox and p22-phox. When stimulated NCF1 interacts with these subunits to form an active enzyme complex that transfers electrons from NADPH to oxygen producing superoxide anion and other ROS.

Pathways

The NCF1/p47-phox component participates mainly in the NADPH oxidase activation pathway and the respiratory burst pathway. NCF1 is connected to the RAC2 GTPase which upon activation stimulates the assembly and activation of the NADPH oxidase complex at the plasma membrane. This pathway is key for the innate immune response and effective clearance of pathogens by phagocytic cells.

Associated diseases and disorders

Mutations or dysfunction in the NCF1/p47-phox gene can lead to chronic granulomatous disease (CGD) which is characterized by a failure to produce ROS and a consequent inability to kill certain bacteria and fungi. Additionally NCF1 has links to autoimmune conditions such as rheumatoid arthritis where impaired ROS production may contribute to pathogenesis. These disease implications also involve other components of the NADPH oxidase complex such as gp91-phox whose dysfunction similarly contributes to these disorders.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

4 product images

  • Flow Cytometry (Intracellular) - Anti-NCF1/p47-phox antibody [EPR27205-231] (ab308256), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-NCF1/p47-phox antibody [EPR27205-231] (ab308256)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HL-1 (mouse atrial muscle cell) (Left panel) compared to RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) (Right panel) cells labelling NCF1/p47-phox with ab308256 at 1/500 dilution (0.1 ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
    A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
    Cells were stained with rabbit IgG or ab308256

  • Immunocytochemistry/ Immunofluorescence - Anti-NCF1/p47-phox antibody [EPR27205-231] (ab308256), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-NCF1/p47-phox antibody [EPR27205-231] (ab308256)

    Immunofluorescent analysis of 80% Methanol-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) cells labelling NCF1/p47-phox with ab308256 at 1/250 dilution (2.004 ug/ml) followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green).

    Confocal image showing cytoplasmic and weak nuclear staining in RAW 264.7 cell line, no staining was observed in HL-1 cell line.

    Low expression control: HL-1.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 ug/ml) (Red). The nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 ug/ml).

  • Western blot - Anti-NCF1/p47-phox antibody [EPR27205-231] (ab308256), expandable thumbnail

    Western blot - Anti-NCF1/p47-phox antibody [EPR27205-231] (ab308256)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Low expression: skeletal muscle, brain.

    In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.

    Exposure time: 180 seconds.

    All lanes: Western blot - Anti-NCF1/p47-phox antibody [EPR27205-231] (ab308256) at 1/1000 dilution

    Lane 1: Mouse lymph node tissue lysate at 20 µg

    Lane 2: Mouse spleen tissue lysate at 20 µg

    Lane 3: Mouse skeletal muscle tissue lysate at 20 µg

    Lane 4: Mouse brain tissue lysate at 20 µg

    Lane 5: Rat spleen tissue lysate at 20 µg

    Lane 6: Rat skeletal muscle tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 44 kDa

    Observed band size: 45 kDa

    Exposure time: 180s

  • Western blot - Anti-NCF1/p47-phox antibody [EPR27205-231] (ab308256), expandable thumbnail

    Western blot - Anti-NCF1/p47-phox antibody [EPR27205-231] (ab308256)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Low expression: HL-1.

    The Lane 3 was developed using a high sensitivity ECL substrate.

    In lanes 1-2, the lysates were stored at -80°C prior to Western Blotting. The bands beneath the target band (32 kDa) are likely to be degradation products. In lane 3, the lysate was freshly made and used for Western Blotting immediately to minimize protein degradation.

    In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.

    Exposure time: 180 seconds.

    All lanes: Western blot - Anti-NCF1/p47-phox antibody [EPR27205-231] (ab308256) at 1/1000 dilution

    Lane 1: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

    Lane 2: HL-1 (mouse atrial muscle cell) whole cell lysate at 20 µg

    Lane 3: RAW 264.7 whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 44 kDa

    Observed band size: 45 kDa

    Exposure time: 180s

Downloads

Product protocols

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

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