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Anti-NCF4 Antibody EP2142Y
Also for NCF4 (NM_013416)
|A synthetic peptide corresponding to residues near the C-terminus of human p40 was used as an immunogen.|
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||WB: 1:25000 - 1:50000; IP: 1:50; IHC-P: 1:100 - 1:250; ICC: 1:100 - 1:250; FC: 1:20 - 1:100
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant (Protein A or G Sepharose)
|Does not react with Mouse, Rat
|Homo sapiens neutrophil cytosolic factor 4, 40kDa (NCF4), transcript variant 2|
|NCF; P40PHOX; SH3PXD4|
|The NADPH oxidase generates superoxide in phagocytic cells. It is important for immunity and its deficiency leads to chronic granulomatous disease (CGD) (1). p40-phox is a newly isolated cytosolic component of the nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase that copurifies with p67-phox. Preliminary evidence indicates that it is a component of the cytosolic complex (2). Maximal activation of NADPH oxidase requires formation of a complex between the p40-phox and p67-phox subunits via association of their PB1 domains. The crystal structure of the p40-phox/p67phox PB1 heterodimer reveals that both domains have a beta grasp topology and that they bind in a front-to-back arrangement through conserved electrostatic interactions between an acidic OPCA motif on p40-phox and basic residues in p67-phox (3). |
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Western blot - NCF4 antibody [EP2142Y]; Anti-NCF4 antibody [EP2142Y] 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - NCF4 antibody [EP2142Y]; ab76158 at 1/100-1/250 dilution staining NCF4 in human spleen by Immunohistochemistry, Paraffin-embedded tissue.
Immunocytochemistry/ Immunofluorescence - Anti-NCF4 antibody [EP2142Y]; ICC/IF image of ab76158 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normalgoat 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 overnight at +4°C. The secondary antibody (green) was , 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 1.43uM.
Flow Cytometry-Anti-NCF4 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 for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed.