NOX2 Antibody - #DF6520

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Product: NOX2 Antibody
Catalog: DF6520
Description: Rabbit polyclonal antibody to NOX2
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Dog
Mol.Wt.: 65kDa; 65kD(Calculated).
Uniprot: P04839
RRID: AB_2838482

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Related Downloads
MS Report
HPLC Report
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
*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,Mouse,Rat
Prediction:
Dog(91%)
Clonality:
Polyclonal
Specificity:
NOX2 Antibody detects endogenous levels of total NOX2.
RRID:
AB_2838482
Cite Format: Affinity Biosciences Cat# DF6520, RRID:AB_2838482.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
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

AMCBX2; CGD; CGD91-phox; CY24B_HUMAN; CYBB; Cytochrome b 245, beta polypeptide; Cytochrome b(558) beta chain; Cytochrome b(558) subunit beta; Cytochrome b-245 heavy chain; Cytochrome b558 subunit beta; GP91 PHOX; gp91-1; gp91-phox; GP91PHOX; Heme-binding membrane glycoprotein gp91phox; NADPH oxidase 2; Neutrophil cytochrome b 91 kDa polypeptide; NOX2; p22 phagocyte B-cytochrome; P91 PHOX; p91-PHOX; Superoxide-generating NADPH oxidase heavy chain subunit;

Immunogens

Immunogen:
Uniprot:

P04839(CY24B_HUMAN) >>Visit HPA database.

Gene(ID):
CYBB(1536)
Expression:
P04839 CY24B_HUMAN:

Detected in neutrophils (at protein level).

Description:
Cytochrome b (-245) is composed of cytochrome b alpha (CYBA) and beta (CYBB) chain. It has been proposed as a primary component of the microbicidal oxidase system of phagocytes. CYBB deficiency is one of five described biochemical defects associated with chronic granulomatous disease (CGD). In this disorder, there is decreased activity of phagocyte NADPH oxidase; neutrophils are able to phagocytize bacteria but cannot kill them in the phagocytic vacuoles. The cause of the killing defect is an inability to increase the cell's respiration and consequent failure to deliver activated oxygen into the phagocytic vacuole. [provided by RefSeq, Jul 2008]
Sequence:
MGNWAVNEGLSIFVILVWLGLNVFLFVWYYRVYDIPPKFFYTRKLLGSALALARAPAACLNFNCMLILLPVCRNLLSFLRGSSACCSTRVRRQLDRNLTFHKMVAWMIALHSAIHTIAHLFNVEWCVNARVNNSDPYSVALSELGDRQNESYLNFARKRIKNPEGGLYLAVTLLAGITGVVITLCLILIITSSTKTIRRSYFEVFWYTHHLFVIFFIGLAIHGAERIVRGQTAESLAVHNITVCEQKISEWGKIKECPIPQFAGNPPMTWKWIVGPMFLYLCERLVRFWRSQQKVVITKVVTHPFKTIELQMKKKGFKMEVGQYIFVKCPKVSKLEWHPFTLTSAPEEDFFSIHIRIVGDWTEGLFNACGCDKQEFQDAWKLPKIAVDGPFGTASEDVFSYEVVMLVGAGIGVTPFASILKSVWYKYCNNATNLKLKKIYFYWLCRDTHAFEWFADLLQLLESQMQERNNAGFLSYNIYLTGWDESQANHFAVHHDEEKDVITGLKQKTLYGRPNWDNEFKTIASQHPNTRIGVFLCGPEALAETLSKQSISNSESGPRGVHFIFNKENF

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Dog
91
Pig
73
Horse
73
Bovine
73
Xenopus
70
Sheep
0
Zebrafish
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P04839 As Substrate

Site PTM Type Enzyme
Y41 Phosphorylation
T42 Phosphorylation
K44 Ubiquitination
N132 N-Glycosylation
N149 N-Glycosylation
Y168 Phosphorylation
N240 N-Glycosylation
K253 Ubiquitination
K294 Ubiquitination
T298 Phosphorylation
Y324 Phosphorylation
K334 Ubiquitination
S486 Phosphorylation
K548 Ubiquitination

Research Backgrounds

Function:

Critical component of the membrane-bound oxidase of phagocytes that generates superoxide. It is the terminal component of a respiratory chain that transfers single electrons from cytoplasmic NADPH across the plasma membrane to molecular oxygen on the exterior. Also functions as a voltage-gated proton channel that mediates the H(+) currents of resting phagocytes. It participates in the regulation of cellular pH and is blocked by zinc.

PTMs:

Glycosylated.

Phosphorylated on Ser and Thr residues.

Undergoes 'Lys-48'-linked polyubiquitination, likely by RNF145, triggering endoplasmic reticulum-associated degradation.

Subcellular Location:

Cell membrane>Multi-pass membrane protein.
Note: As unassembled monomer may localize to the endoplasmic reticulum.

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 neutrophils (at protein level).

Subunit Structure:

Composed of a heavy chain (beta) and a light chain (alpha). 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 with NCF1. Interacts with calprotectin (S100A8/9). Interacts with NRROS; the interaction is direct and impairs formation of a stable NADPH oxidase complex. Interacts with CYBC1; CYBC1 may act as a chaperone stabilizing Cytochrome b-245 heterodimer.

Research Fields

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

· Cellular Processes > Cell growth and death > Ferroptosis.   (View pathway)

· Cellular Processes > Cell growth and death > Necroptosis.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Human Diseases > Infectious diseases: Parasitic > Leishmaniasis.

· Organismal Systems > Immune system > NOD-like receptor signaling pathway.   (View pathway)

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

References

1). Oxidative stress induced by NOX2 contributes to neuropathic pain via plasma membrane translocation of PKCε in rat dorsal root ganglion neurons. Journal of Neuroinflammation (PubMed: 33952299) [IF=9.3]

Application: WB    Species: Rat    Sample: DRGs

Fig. 1. SNI induces NOX2 upregulation in DRG neurons. a Paw withdrawal threshold (PWT) after SNI (n = 5–6/group). Mann–Whitney U test. *p < 0.05, **p < 0.01 versus the sham group. b NOX2 expression in L4–L6 DRGs from rats in (a) at 1, 7, and 14 days after SNI and sham surgery was determined by Western blotting (n = 4/group). n.s. indicates not significant. One-way ANOVA followed by Tukey’s test. *p < 0.05 versus the sham group. c Representative double-immunofluorescence staining showing the colocalization of NOX2 with IB4 (c(a)), CGRP (c(b)), NF-200 (c(c)), and CD11b (c(d)) but not GFAP (c(e)) at day 7 after SNI. c(f) The histogram shows the percentage of NOX2-positive cells among the various cell markers (n = 6/group)

Application: IF/ICC    Species: Rat    Sample: DRGs

Fig. 1. SNI induces NOX2 upregulation in DRG neurons. a Paw withdrawal threshold (PWT) after SNI (n = 5–6/group). Mann–Whitney U test. *p < 0.05, **p < 0.01 versus the sham group. b NOX2 expression in L4–L6 DRGs from rats in (a) at 1, 7, and 14 days after SNI and sham surgery was determined by Western blotting (n = 4/group). n.s. indicates not significant. One-way ANOVA followed by Tukey’s test. *p < 0.05 versus the sham group. c Representative double-immunofluorescence staining showing the colocalization of NOX2 with IB4 (c(a)), CGRP (c(b)), NF-200 (c(c)), and CD11b (c(d)) but not GFAP (c(e)) at day 7 after SNI. c(f) The histogram shows the percentage of NOX2-positive cells among the various cell markers (n = 6/group)
2). Kaempferol alleviates calcium oxalate crystal-induced renal injury and crystal deposition via regulation of the AR/NOX2 signaling pathway. PHYTOMEDICINE (PubMed: 33852977) [IF=7.9]

Application: IHC    Species: Human    Sample: HK-2 cells

Fig. 3. Kaempferol inhibited renal oxidative stress and inflammation in vivo. (a-e) NOX2, iNOS, NFκB-p65 and F4/80 stainings of paraffin embedded kidney sections were evaluated. (f) ROS level in renal tissue was detected by fluorescence microplate after loading with DCFH-DA. Moreover, (g) H2O2 expression, as well as (h) GSH, SOD and MDA levels were detected in mice renal tissues. (i-n) The mRNA levels of IL-1β, IL-6, TNF-α, IL-10, IL-4 and Arg1 in renal tissues were measured by RT-qPCR. (o) Group annotations were applied for all histograms. Data were expressed as the fold changes of the experimental group to the NC group, and were represented as means ± SD. * p < 0.05 vs. NC group; ** p < 0.05 as GA + Kae (25 mg/kg) vs. GA group, GA + Kae (50 mg/kg) vs. GA group, and GA + Kae (25 mg/kg) vs. GA + Kae (50 mg/kg) group, respectively.
3). Bi-phasic effect of gelatin in myogenesis and skeletal muscle regeneration. Disease Models & Mechanisms (PubMed: 34821368) [IF=4.3]

Application: WB    Species: Mouse    Sample:

Fig. 4.Linear and bell-shaped dose response of reactive oxygen species (ROS) and the antioxidant system to gelatin. (A,B) Flow cytometry analysis (A) and quantification (B) of cells stained with 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA; DCF) (ROS indicator). Count (y-axis in A) indicates cell counts (n=3). (C) Production of specific ROS: O2−, ·OH and H2O2 (n=3). (D) Activities of antioxidases: superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and catalase (CAT) (n=3). (E,F) Western blots (E) and quantification (F) of NOX2 and NOX4 in the cells cultured on gelatin-coated dishes at 90% confluence (n=3). β-Actin is used as a loading control. (G) Quantification of MitoSOX+ cells obtained by flow cytometry (n=3). (H) Flow cytometry analysis of the cells treated with diphenyliodonium (DPI; 0.5 μM, NOX2 inhibitor) and rotenone (ROT; 10 μM, an inhibitor of mitochondrial respiratory complex I) (n=3). Significance was determined by unpaired two-tailed Student's t-test with Welch's correction (B-D,F-G) and one-way ANOVA with Tukey's post-hoc test (H).
4). Eupatilin attenuates doxorubicin-induced cardiotoxicity by activating the PI3K-AKT signaling pathway in mice. Molecular and Cellular Biochemistry (PubMed: 37222879) [IF=4.3]
5). Diosmin mitigates high glucose-induced endoplasmic reticulum stress through PI3K/AKT pathway in HK-2 cells. BMC Complementary Medicine and Therapies (PubMed: 35477428) [IF=3.9]

Application: WB    Species: human    Sample: HK-2 cells

Fig. 6 |The efect of diosmin on oxidative stress-, infammation-, apoptosis-, and autophagy-related factors in high glucose-mediated HK-2 cells.A-J. The efect of diosmin on oxidative stress-, infammation-, apoptosis-, and autophagy-related factors in high glucose-mediated HK-2 cells was
6). Idebenone attenuates cerebral inflammatory injury in ischemia and reperfusion via dampening NLRP3 inflammasome activity. MOLECULAR IMMUNOLOGY (PubMed: 32438202) [IF=3.6]

Application: WB    Species: mouse    Sample: BV3 cells

Fig. 3. |Idebenone inhibited NLRP3 signaling in BV2 cells under OGD/R conditions. (a) Immunoblot analysis of NQO1, NQO2, NOX2 and NLRP3 signaling in BV2 cells with or without treatment of idebenone after OGD/R.
7). Lycopene Exerts Renoprotective Effects Through Inhibiting Pyroptosis Induced by Hyperoxaluria in Vivo and in Vitro.
8). Ficus pandurata Hance Inhibits Ulcerative Colitis and Colitis-Associated Secondary Liver Damage of Mice by Enhancing Antioxidation Activity. Oxidative Medicine and Cellular Longevity (PubMed: 34966476)

Application: WB    Species: Mouse    Sample:

Figure 5 FPH enhances the colonic antioxidation in the UC mice induced by DSS. Effect of FPH on the levels of MDA (a), T-SOD (b), and GSH-Px (c) in the colonic homogenate detected by ELISA. (d) Effect of FPH on the protein expressions of Keap1, Nrf2, HO1, NQO1, p22-phox, and NOX2 in colon tissues detected by Western blot. Results were expressed as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01 vs. the control group; #p < 0.05, ##p < 0.01 vs. the model group (DSS only).

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