NAD(P)H Dehydrogenase, Quinone 1 (NQO1) antibody

Details for Product No. ABIN268227
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Antigen
Synonyms zgc:77191, wu:fb63c10, nqo1, DHQU, DIA4, DTD, NMOR1, NMORI, QR1, Dia4, AV001255, Dtd, Nmo-1, Nmo1, Nmor1, Ox-1, Ox1, Qr1, NADPH-d
Reactivity
Dog (Canine), Human, Monkey, Rat (Rattus)
(107), (29), (28), (15), (12), (9), (4), (1)
Host
Mouse
(59), (39), (10), (1)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(4), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Application
Western Blotting (WB), Flow Cytometry (FACS), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP)
(99), (32), (29), (27), (25), (16), (14), (10), (6), (6), (3), (2), (2), (1), (1)
Pubmed 5 references available
Quantity 0.025 mL
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Catalog No. ABIN268227
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Immunogen Full length recombinant NQO1 from human lung
Clone A180
Isotype IgG1
Specificity Does not cross-react with NQO2.
Cross-Reactivity (Details) Mouse reactivity reported in scientific literature (PMID: 24475200)
Purification Protein G purified
Alternative Name NQO1
Background NQO1 is a 2-electron reductase that detoxifies quinones derived from the oxidation ofphenolic metabolites of benzene. Individuals homozygous for the T/T form of the609C-T polymorphism have an increased risk of benzene hematotoxicity. Alternate Names: anti-DHQU antibody, anti-DIA4 antibody, anti-Diaphorase (NADH/NADPH) (cytochromeb-5 reductase) antibody, anti-Diaphorase (NADH/NADPH) antibody, anti-Diaphorase 4antibody, anti-DTD antibody, anti-NAD(P)H dehydrogenase quinone 1 antibody,anti-NAD(P)H: menadione oxidoreductase 1 dioxin inducible 1 antibody, anti-NMOR1antibody, anti-NMORI antibody, anti-QR1 antibody, anti-NQO-1 antibody, anti-NADPHquinone oxidoreductase-1 antibody.
Gene Symbol: NQO1
Gene ID 1728
UniProt P15559
Research Area Signaling, Metabolism
Application Notes This NQO1 Antibody (A180) is useful for Western blot, Immunoprecipitation, Immunocytochemistry/Immunofluorescence, Immunohistochemistry on frozen and paraffin-embedded sections and Flow Cytometry. In Western blot a band can be seen at ~ 31 kDa representing NQO1. In ICC/IF, cytoplasmic staining was observed in U2OS cells. In IHC-P, staining was observed in the cytoplasm of human breast cancer tissue. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.
Recommended dilutions: Flow Cytometry 1:10-1:1000, Immunocytochemistry/Immunofluorescence 1:50, Immunohistochemistry 1:100, Immunohistochemistry-Frozen 1:100, Immunohistochemistry-Paraffin 1:100, Immunoprecipitation 1:1000, Western Blot 1:1000
Protocol Protocol specific for NQO1 Antibody Western Blot Protocol
1. Perform SDS-PAGE on samples to be analyzed, loading 40 µg of total protein per lane.
. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
. Rinse the blot.
. Block the membrane using standard blocking buffer for at least 1 hour.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturers instructions.Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %.Immunohistochemistry-Paraffin Embedded SectionsAntigen Unmasking:Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.
0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes. Staining:
. Wash sections in deionized water three times for 5 minutes each.
. Wash sections in wash buffer for 5 minutes.
. Block each section with 100-400 µL blocking solution for 1 hour at room temperature.
. Remove blocking solution and add 100-400 µL diluted primary antibody. Incubate overnight at 4 C.
. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
. Add 100-400 µL biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
. Add 100-400 µL Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
. Wash sections three times in wash buffer for 5 minutes each.
. Add 100-400 µL DAB substrate to each section and monitor staining closely.
. As soon as the sections develop, immerse slides in deionized water.
. Counterstain sections in hematoxylin.
. Wash sections in deionized water two times for 5 minutes each.
. Dehydrate sections.
. Mount coverslips.*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
Restrictions For Research Use only
Format Liquid
Concentration 1.0 mg/mL
Buffer Tris-glycine, 150 mM NaCl, Sodium Azide
Preservative Sodium azide
Precaution of Use WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handling Advice Avoid freeze-thaw cycles
Storage 4 °C
Storage Comment 4 °C short term. Aliquot and store at -20 °C long term.
General Siegel, Franklin, Ross: "Immunohistochemical detection of NAD(P)H:quinone oxidoreductase in human lung and lung tumors." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 4, Issue 9, pp. 2065-70, 1998 (PubMed).

Slitt, Cherrington, Dieter et al.: "trans-Stilbene oxide induces expression of genes involved in metabolism and transport in mouse liver via CAR and Nrf2 transcription factors." in: Molecular pharmacology, Vol. 69, Issue 5, pp. 1554-63, 2006 (PubMed).

Lee, Bhora, Sun et al.: "Dietary flaxseed enhances antioxidant defenses and is protective in a mouse model of lung ischemia-reperfusion injury." in: American journal of physiology. Lung cellular and molecular physiology, Vol. 294, Issue 2, pp. L255-65, 2008 (PubMed).

Cho, Imani, Miller-DeGraff et al.: "Antiviral activity of Nrf2 in a murine model of respiratory syncytial virus disease." in: American journal of respiratory and critical care medicine, Vol. 179, Issue 2, pp. 138-50, 2009 (PubMed).

Salama, Kamel, Diaz-Arrastia et al.: "Effect of tumor necrosis factor-alpha on estrogen metabolism and endometrial cells: potential physiological and pathological relevance." in: The Journal of clinical endocrinology and metabolism, Vol. 94, Issue 1, pp. 285-93, 2009 (PubMed).

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