Phospho-NCF1/p47-phox (Ser359) Antibody - #AF3167

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Product: Phospho-NCF1/p47-phox (Ser359) Antibody
Catalog: AF3167
Description: Rabbit polyclonal antibody to Phospho-NCF1/p47-phox (Ser359)
Application: WB IHC IF/ICC
Reactivity: Human, Rat
Mol.Wt.: 45kDa; 45kD(Calculated).
Uniprot: P14598
RRID: AB_2834599

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MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Rat
Clonality:
Polyclonal
Specificity:
Phospho-NCF1/p47-phox (Ser359) Antibody detects endogenous levels of NCF1/p47-phox only when phosphorylated at Serine 359.
RRID:
AB_2834599
Cite Format: Affinity Biosciences Cat# AF3167, RRID:AB_2834599.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

47 kDa autosomal chronic granulomatous disease protein; 47 kDa neutrophil oxidase factor; NADPH oxidase organizer 2; NCF 47K; NCF-1; NCF-47K; Ncf1; NCF1_HUMAN; Neutrophil cytosol factor 1; Neutrophil cytosolic factor 1; neutrophil cytosolic factor 1, (chronic granulomatous disease, autosomal 1); Neutrophil NADPH oxidase factor 1; Nox organizer 2; Nox organizing protein 2; Nox-organizing protein 2; NOXO2; p47 phox; p47-phox; SH3 and PX domain containing protein 1A; SH3 and PX domain-containing protein 1A; SH3PXD1A;

Immunogens

Immunogen:
Uniprot:

P14598(NCF1_HUMAN) >>Visit HPA database.

Gene(ID):
NCF1(653361)
Expression:
P14598 NCF1_HUMAN:

Detected in peripheral blood monocytes and neutrophils (at protein level).

Description:
The 47-kilodalton cytosolic subunit of the multi-protein complex known as NADPH oxidase is found in neutrophils. The holo-oxidase produces a burst of superoxide which is delivered to the lumen of the neutrophil phagosome. Contains 2 SH2 domains.
Sequence:
MGDTFIRHIALLGFEKRFVPSQHYVYMFLVKWQDLSEKVVYRRFTEIYEFHKTLKEMFPIEAGAINPENRIIPHLPAPKWFDGQRAAENRQGTLTEYCSTLMSLPTKISRCPHLLDFFKVRPDDLKLPTDNQTKKPETYLMPKDGKSTATDITGPIILQTYRAIANYEKTSGSEMALSTGDVVEVVEKSESGWWFCQMKAKRGWIPASFLEPLDSPDETEDPEPNYAGEPYVAIKAYTAVEGDEVSLLEGEAVEVIHKLLDGWWVIRKDDVTGYFPSMYLQKSGQDVSQAQRQIKRGAPPRRSSIRNAHSIHQRSRKRLSQDAYRRNSVRFLQQRRRQARPGPQSPGSPLEEERQTQRSKPQPAVPPRPSADLILNRCSESTKRKLASAV

PTMs - P14598 As Substrate

Site PTM Type Enzyme
Y41 Phosphorylation
T45 Phosphorylation
Y48 Phosphorylation
T133 Phosphorylation Q9NWZ3 (IRAK4)
T138 Phosphorylation Q9NWZ3 (IRAK4)
Y139 Phosphorylation
T153 Phosphorylation Q9NWZ3 (IRAK4)
S208 Phosphorylation P68400 (CSNK2A1)
Y279 Phosphorylation
S283 Phosphorylation P68400 (CSNK2A1)
S288 Phosphorylation Q9NWZ3 (IRAK4)
S303 Phosphorylation P17252 (PRKCA) , Q05513 (PRKCZ) , Q05655 (PRKCD) , P05771 (PRKCB)
S304 Phosphorylation Q05513 (PRKCZ) , P05771-2 (PRKCB) , P17252 (PRKCA) , P31749 (AKT1) , Q05655 (PRKCD)
S310 Phosphorylation
S315 Phosphorylation P05771 (PRKCB) , P17252 (PRKCA) , Q05513 (PRKCZ) , Q05655 (PRKCD)
S320 Phosphorylation Q9NWZ3 (IRAK4) , P17612 (PRKACA) , P05771-2 (PRKCB) , Q05513 (PRKCZ) , Q05655 (PRKCD) , P17252 (PRKCA)
S328 Phosphorylation Q05655 (PRKCD) , P31749 (AKT1) , P17252 (PRKCA) , P17612 (PRKACA) , Q05513 (PRKCZ) , P05771 (PRKCB)
S345 Phosphorylation Q16539 (MAPK14) , Q9NWZ3 (IRAK4) , P28482 (MAPK1) , P27361 (MAPK3)
S348 Phosphorylation P68400 (CSNK2A1) , P28482 (MAPK1) , P27361 (MAPK3) , Q9NWZ3 (IRAK4) , Q16539 (MAPK14)
T356 Phosphorylation Q9NWZ3 (IRAK4)
S359 Phosphorylation Q9NWZ3 (IRAK4) , Q05513 (PRKCZ) , P05771-2 (PRKCB) , P17252 (PRKCA) , Q05655 (PRKCD) , P17612 (PRKACA)
S370 Phosphorylation Q05513 (PRKCZ) , Q9NWZ3 (IRAK4) , P05771 (PRKCB) , Q05655 (PRKCD) , P17612 (PRKACA) , P17252 (PRKCA)
S379 Phosphorylation Q05513 (PRKCZ) , Q05655 (PRKCD) , P05771 (PRKCB) , P17252 (PRKCA)
S381 Phosphorylation
T382 Phosphorylation

Research Backgrounds

Function:

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

PTMs:

Phosphorylated by PRKCD; phosphorylation induces activation of NCF1 and NADPH oxidase activity.

Subcellular Location:

Cytoplasm>Cytosol. Membrane>Peripheral membrane protein>Cytoplasmic side.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Detected in peripheral blood monocytes and neutrophils (at protein level).

Subunit Structure:

Component of an NADPH oxidase complex composed of a heterodimer formed by the membrane proteins CYBA and CYBB and the cytosolic subunits NCF1, NCF2 and NCF4. Interacts (via C-terminus) with NCF2 (via the C-terminal SH3 domain). Interacts with NCF4. Interacts with CYBB. Interacts (via the second SH3 domain) with CYBA. Interacts with NOXA1. Interacts with ADAM15. Interacts with TRAF4. Interacts with FASLG. Interacts with PARK7 (via C-terminus); the interaction is enhanced by LPS and modulates NCF1 phosphorylation and membrane translocation (By similarity).

Family&Domains:

The PX domain mediates interaction with phosphatidylinositol 3,4-bisphosphate and other anionic phospholipids. In the autoinhibited, unphosphorylated state an intramolecular interaction with the C-terminal SH3 domain precludes phospholipid binding and interaction with CYBA. Phosphorylation disrupts the autoinhibited state.

Research Fields

· Cellular Processes > Transport and catabolism > Phagosome.   (View pathway)

· Human Diseases > Infectious diseases: Parasitic > Leishmaniasis.

· Organismal Systems > Immune system > Chemokine signaling pathway.   (View pathway)

· Organismal Systems > Development > Osteoclast differentiation.   (View pathway)

· Organismal Systems > Immune system > Fc gamma R-mediated phagocytosis.   (View pathway)

· Organismal Systems > Immune system > Leukocyte transendothelial migration.   (View pathway)

References

1). Oxidation of Reduced Graphene Oxide via Cellular Redox Signaling Modulates Actin-Mediated Neurotransmission. ACS Nano, 2020 (PubMed: 32057235) [IF=17.1]

Application: WB    Species: rat    Sample: PC12

Figure S6 Oxidized rGO increased lipid peroxidation and NOX2 activation in PC12 cells. Oxidized rGO (c-rGO) was collected after pristine rGO (p-rGO) was cultured with PC12 cells for 3 h. (A-B) PC12 cells were pretreated with NAC for 30 min before being treated with two kinds of rGO (20 µg/mL) for 1 h and stained with BODIPY 581/591 C11 reagent. The lipid peroxidation degrees were quantified via FITC fluorescence intensity, with n = 10 fields per experimental condition from 3 independent tests. Scale bar: 100 μm. (C-D) After PC12 cells were treated as described in (A), plasma membrane proteins were isolated to detect the expression levels of NOX2, p-p47phox, p-p67phox and Rac2. The protein levels were quantified after normalization to ATPase. The results represent the mean ± SD of three independent experiments. *p<0.05, **p<0.01, ***p<0.001 compared with the control group and with the p-rGO group.
2). Preclinical efficacy of TZG in myofascial pain syndrome by impairing PI3K-RAC2 signaling-mediated neutrophil extracellular traps. iScience, 2023 (PubMed: 37860777) [IF=5.8]

Application: WB    Species: Rat    Sample:

Figure 5 TZG significantly regulated the expression of NETs formation-related indicators, and the corresponding proteins in PI3K-RAC2 signaling (A–E). Serum levels of dsDNA, IL-8, TNF-α, CitH3, and MPO detected by ELISA analyses, respectively. Samples in (A)∼(E) were obtained from the blood supernatant of rats in different groups. (F–I) The ratios of p-PI3K/PI3K and p-P47/P47, and the expression levels of RAC2 and PAD4 proteins detected by western blot analyses, respectively. Samples in (F)∼(I) were obtained from MTrPs tissues of rats in different groups. The number of rats in each group was 8. Data are expressed as the mean ± S.D. “∗,” “∗∗,” and “∗∗∗,” p < 0.05, p < 0.01, and p < 0.001, respectively, comparison with the normal control group. “#,” “##,” and “###,” p < 0.05, p < 0.01, and p < 0.001, respectively, comparison with the MPS model group.
3). CHANGES IN THE RAGE/NADPH OXIDASE SIGNALING PATHWAY AND OXIDATIVE STRESS LEVELS IN SH-SY5Y CELLS EXPOSED TO HIGH LEVELS OF FLUORIDE.. Fluoride, 2022 [IF=0.9]

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