Overview

  • Product name
    Anti-NCF4 antibody [EP2142Y]
    See all NCF4 primary antibodies
  • Description
    Rabbit monoclonal [EP2142Y] to NCF4
  • Tested applications
    Suitable for: ICC/IF, WB, IP, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    A synthetic peptide corresponding to residues near the C terminus of human NCF4

  • Positive control
    • HL60 cell lysate; human spleen tissue. IF/ICC: MCF7 cell line.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

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

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

Properties

Applications

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
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.
Flow Cyt 1/20 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • 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
  • Database links
  • Alternative names
    • CGD3 antibody
    • MGC3810 antibody
    • NCF 4 antibody
    • NCF antibody
    • NCF-4 antibody
    • Ncf4 antibody
    • NCF4_HUMAN antibody
    • Neutrophil cytosol factor 4 antibody
    • Neutrophil cytosolic factor 4 antibody
    • Neutrophil NADPH oxidase factor 4 antibody
    • p40-phox antibody
    • p40phox antibody
    • SH3 and PX domain-containing protein 4 antibody
    • SH3PXD4 antibody
    see all

Images

  • 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.

  • ab76158 at 1/100-1/250 dilution staining NCF4 in human spleen by Immunohistochemistry, Paraffin-embedded tissue.
  • Anti-NCF4 antibody [EP2142Y] (ab76158) at 1/50000 dilution + HL60 cell lysate at 10 µg

    Secondary
    goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size : 39 kDa
    Observed band size : 39 kDa
  • 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 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:
  • 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). Read more (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 . Read more (PubMed: 22235113) »

See all 4 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (monocyte-derived macrophage)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
30 µg
Specification
monocyte-derived macrophage
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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Verified customer

Submitted May 25 2015

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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