p22phox Antibody [G20B7] | Primary Antibodies

Application: Reactivity: Human, Mouse, Rat

Usage Information

Dilution
WB1:1000 IHC1:500 - 1:2000 IF1:800 - 1:3200

Application
WB, IHC, IF

Reactivity
Human, Mouse, Rat

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
22 kDa

Positive Control	Mouse small intestine; Mouse spleen; Rat liver; Human hepatocellular carcinoma; Human hepatocellular carcinoma; Human small intestine; Human spleen; Human brain; MC-38 syngeneic tumor; RPMI 8226 cells; RAW 264.7 cells; THP-1 cells; HCC 1937 cells; IGROV-1 cells
Negative Control	A-172 cells; RH-30 cells

Datasheet & SDS

Biological Description

Specificity
p22phox Antibody [G20B7] detects endogenous levels of total p22phox protein.

Clone
G20B7

Synonym(s)
Cytochrome b-245 light chain; Cytochrome b(558) alpha chain; p22 phagocyte B-cytochrome; p22-phox (p22phox); CYBA

Background
p22phox, also known as CYBA, is a critical membrane-spanning subunit of the NADPH oxidase (NOX) complex. It forms a stable heterodimer with the catalytic NOX2 (gp91phox) subunit in phagocytes and associates with NOX1, NOX3, and NOX4 in non-phagocytic cells to facilitate the production of superoxide, a process essential for innate immunity and redox signaling. p22phox features a compact four-helix bundle transmembrane domain composed of helices M1 to M4, which laterally interacts with the transmembrane domain of NOX2 across three conserved interfaces involving NOX2’s M3, M4, and M5 and p22phox’s M1 and M4. Its extracellular loops form a glycan shield, while the cytoplasmic C-terminus, containing the Thr147 phosphorylation site, extends into the cytosol to provide docking sites for cytosolic factors. The segment spanning residues 128 to 136 inserts into a crevice in NOX2, stabilizing the overall assembly. p22phox stabilizes the latent NOX complex and facilitates its activation: upon phagocytic stimulation, cytosolic subunits such as p47phox, p67phox, p40phox, and the small GTPase Rac translocate to the membrane, initiating conformational changes that allow docking of NOX2’s dehydrogenase domain. This enables the oxidation of NADPH, electron transfer through flavin and heme cofactors, and reduction of molecular oxygen to superoxide, driving microbicidal reactive oxygen species (ROS) bursts and regulating vascular smooth muscle growth, inflammation, and tumorigenesis. p22phox is indispensable for phagosome maturation, pathogen killing, and also plays non-immune roles such as maintaining endothelial function. Phosphorylation at Thr147 enhances complex assembly. Mutations in p22phox cause chronic granulomatous disease due to impaired ROS production and recurrent infections, whereas overexpression is linked to ROS-driven pathologies such as cancer, where it stabilizes HIF-2α via tuberin phosphorylation, atherosclerosis, neurodegeneration (as seen in SOD1-linked ALS and amyloid-beta-exacerbated neuronal death in Alzheimer’s disease), hyperglycemia, and NF-κB-mediated smooth muscle proliferation.

Background

p22phox, also known as CYBA, is a critical membrane-spanning subunit of the NADPH oxidase (NOX) complex. It forms a stable heterodimer with the catalytic NOX2 (gp91phox) subunit in phagocytes and associates with NOX1, NOX3, and NOX4 in non-phagocytic cells to facilitate the production of superoxide, a process essential for innate immunity and redox signaling. p22phox features a compact four-helix bundle transmembrane domain composed of helices M1 to M4, which laterally interacts with the transmembrane domain of NOX2 across three conserved interfaces involving NOX2’s M3, M4, and M5 and p22phox’s M1 and M4. Its extracellular loops form a glycan shield, while the cytoplasmic C-terminus, containing the Thr147 phosphorylation site, extends into the cytosol to provide docking sites for cytosolic factors. The segment spanning residues 128 to 136 inserts into a crevice in NOX2, stabilizing the overall assembly. p22phox stabilizes the latent NOX complex and facilitates its activation: upon phagocytic stimulation, cytosolic subunits such as p47phox, p67phox, p40phox, and the small GTPase Rac translocate to the membrane, initiating conformational changes that allow docking of NOX2’s dehydrogenase domain. This enables the oxidation of NADPH, electron transfer through flavin and heme cofactors, and reduction of molecular oxygen to superoxide, driving microbicidal reactive oxygen species (ROS) bursts and regulating vascular smooth muscle growth, inflammation, and tumorigenesis. p22phox is indispensable for phagosome maturation, pathogen killing, and also plays non-immune roles such as maintaining endothelial function. Phosphorylation at Thr147 enhances complex assembly. Mutations in p22phox cause chronic granulomatous disease due to impaired ROS production and recurrent infections, whereas overexpression is linked to ROS-driven pathologies such as cancer, where it stabilizes HIF-2α via tuberin phosphorylation, atherosclerosis, neurodegeneration (as seen in SOD1-linked ALS and amyloid-beta-exacerbated neuronal death in Alzheimer’s disease), hyperglycemia, and NF-κB-mediated smooth muscle proliferation.

References
https://pubmed.ncbi.nlm.nih.gov/25263488/ https://pubmed.ncbi.nlm.nih.gov/36413210/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%