Immunofluorescent analysis of Nox3 (Violet) in mouse bone marrow cells (Up). The cells were fixed with 2% paraformaldehyde in PBS for 30 minutes at RT, permeabilized with 0.5% Triton X-100 in PBS for 30 minutes at RT, and blocked with blocking buffer for 4 hours in cold room. Cells were stained with rabbit polyclonal anti-Nox3 antibody (Product # PA5-38036) at a dilution of 1:200 in blocking buffer overnight in cold room, wash 5 times with 1 xPBS, incubated with goat anti-rabbit IgG antibody conjugated with Cy5 (Product # A10523) at a dilution of 1:1000 for 1 hours at RT (panel b: Violet) and wash 5 times with 1xPBS before overlaid with cover slip. Nuclei (panel a: blue) were stained with DAPI (Product # D1306) during the blocking step. Panels a and b are a merged image and magnified squire area, respectively. Images were taken on a Nikon A1 confocal microscopy at 60x Objective lens with oil used. Data courtesy of Dr. Zhi Wen from the University of Wisconsin, Madison.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||15 amino acid peptide near the amino terminus of human NOX3|
|Purification||Antigen affinity chromatography|
|Contains||0.02% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 3 months. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||5 µg/ml|
|Western Blot (WB)||1-2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA5-38036 detects Nox3 in mouse bone marrow and humanG-CSF mobilized CD34+ cells in immunofluorescence analysis.
A recommended positive control is 293 cell lysate.
NOX3 is not expected to cross-react with other NOX proteins.
The NOX family of NAPDH oxidases is comprised of seven transmembrane proteins that oxidize intracellular NAPDH/NADH, causing electron transport across the membrane and the reduction of molecular oxygen to superoxide. NOX3 is expressed predominantly in the inner ear and is involved in the biogenesis of otoconia/otolith. It has been suggested that NOX3 is activated by the Transient Receptor Potential Vanilloid 1 (TRPV1), and this activity causes increased levels of reactive oxygen species in the inner ear, which in turn leads to STAT1-mediated inflammation and hearing loss.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.