NQO-1 Antibody (A180) - Azide and BSA Free

Western Blot: NQO-1 Antibody (A180) - Azide and BSA Free [NBP2-80885] - Detection of NQO1 in A549 lysates using NB 200-209. Lane 1: 0.5 ug/ml. Lane 2: 2 ug/ml. Exposure: 1 second. Image from the standard format of this antibody.

Immunocytochemistry/Immunofluorescence: NQO-1 Antibody (A180) - Azide and BSA Free [NBP2-80885] - NQO1 antibody was tested in U2OS cells with Dylight 488 (green). Nuclei and beta-tubulin were counterstained with DAPI (blue) and Dylight 550 (red). Image from the standard format of this antibody.

Immunohistochemistry-Paraffin: NQO-1 Antibody (A180) - Azide and BSA Free [NBP2-80885] - Immunohistochemistry showing Anti-NQ01 on formalin fixed paraffin embedded human kidney tissue Image from the standard format of this antibody.

Flow Cytometry: NQO-1 Antibody (A180) - Azide and BSA Free [NBP2-80885] - Detection of NQO-1 in Human A549 Cell Line by Flow Cytometry. Human A549 cell line was stained with Mouse Anti- NQO-1 Monoclonal Antibody (Catalog # NB200-209, filled histogram), or Mouse IgG1 isotype control (Catalog # MAB002, open histogram) followed by APC-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). Images may not be copied, printed or otherwise disseminated without express written permission of Novus Biologicals a Bio-Techne brand. Image from the standard format of this antibody.

Western Blot: NQO-1 Antibody (A180) - Azide and BSA Free [NBP2-80885] - Antioxidant enzyme synthesis in response to orange oil treatment. Immunoblot analysis demonstrating expression of NQO1 protein at 6, 12 and 24 hrs following 15 min treatment of BEAS-2B cells with the oil preparation or time-matched soy oil control. Representative blots from one of three separate experiments are shown above. Densitometric evaluations of each target protein blot normalized to its corresponding GAPDH for all three experiments are provided below. Bars represent mean +/- SEM. Image collected and cropped by CiteAb from the following publication (//respiratory-research.biomedcentral.com/articles/10.1186/1465-9921-12-92), licensed under a CC-BY license. Image from the standard format of this antibody.

Immunohistochemistry: NQO-1 Antibody (A180) - Azide and BSA Free [NBP2-80885] - Analysis of NQO1 in human breast cancer using DAB with hematoxylin counterstain. Image from the standard format of this antibody.

Flow Cytometry: NQO-1 Antibody (A180) - Azide and BSA Free [NBP2-80885] - An intracellular stain was performed on U-87 cells with NQO-1 [A180] Antibody NB200-209AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were directly conjugated to Alexa Fluor 647.

Flow Cytometry: NQO-1 Antibody (A180) - Azide and BSA Free [NBP2-80885] - An intracellular stain was performed on U-87 cells with NQO-1 [A180] Antibody NB200-209C (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were directly conjugated to DyLight 650.

Simple Western: NQO-1 Antibody (A180) - Azide and BSA Free [NBP2-80885] - Simple Western lane view shows a specific band for NQO-1 in 0.5 mg/ml of A431 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. Image from the standard format of this antibody.

• Reactivity: Hu, Mu, Rt, Ca, Pm
• Applications: WB, Simple Western, Flow, ICC/IF, IHC, IP
• Clone: A180
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: Azide and BSA Free
• Concentration: 1 mg/ml

NQO-1 Antibody (A180) - Azide and BSA Free Summary

• Description: Novus Biologicals Mouse NQO-1 Antibody (A180) - Azide and BSA Free (NB200-209) is a monoclonal antibody validated for use in IHC, WB, Flow, ICC/IF, Simple Western and IP. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: Full length recombinant NQO1 from human lung [UniProt# P15559]
• Localization: Cytoplasmic (primarily), Nuclear (faintly)
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: NQO1
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Flow (Intracellular)
  • Flow Cytometry 1-2 ug/ml
  • Immunocytochemistry/ Immunofluorescence 1:50
  • Immunohistochemistry 1:100
  • Immunohistochemistry-Frozen 1:100
  • Immunohistochemistry-Paraffin 1:100
  • Immunoprecipitation 1:1000
  • Simple Western 1:100
  • Western Blot 1:1000
• Application Notes: In Western blot a band can be seen at approx. 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.
  The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
  See Simple Western Antibody Database for Simple Western validation: antibody dilution of 1:100. Separated by Size-Wes, Sally Sue/Peggy Sue.
  The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Theoretical MW: 31 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 24475200)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Tris-Glycine, 0.15 M NaCl
• Preservative: No Preservative
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for NQO-1 Antibody (A180) - Azide and BSA Free

• Azoreductase
• DHQU
• DIA4
• DIA4azoreductase
• diaphorase (NADH/NADPH) (cytochrome b-5 reductase)
• Diaphorase-4
• Dioxin-inducible 1
• DTD
• DTDEC 1.6.5.2
• DT-diaphorase
• Menadione Reductase
• NAD(P)H dehydrogenase [quinone] 1
• NAD(P)H dehydrogenase, quinone 1
• NAD(P)H:menadione oxidoreductase 1
• NAD(P)H:Quinone acceptor oxidoreductase type 1
• NAD(P)H:quinone oxidoreductase 1
• NAD(P)H:quinone oxireductase
• NMOR1
• NMOR1diaphorase-4
• NQO1
• NQO-1
• Phylloquinone reductase
• QR1
• QR1NMORI
• Quinone Reductase 1

Background

NAD(P)H:quinone oxidoreductase (NQO) is a ubiquitous flavoprotein enzyme which catalyzes the two-electron reduction/detoxification of quinones/quinones derivatives and it has two members - NQO1 (alias DT-diaphorase) which uses NADPH as a source of electrons, and NQO2 (NRH:quinone oxidoreductase 2) which employs dihydronicotinamide riboside (NRH) as electron donor. Expressed as cytosolic enzyme with maximum levels in liver, NQO1 is highly inducible by oxidants, electrophiles, phenolic compounds etc and its expression is regulated by Keap1/Nrf2/ARE pathway in mammals. NQO1 gene promotor contains multiple AREs (antioxidant response elements) and a functional XRE (xenobiotic response element), and AhR binding to XRE is critical for NQO1 induction by xenobiotics. Moreover, Nrf2/ARE and AhR/XRE have been proposed to coordinate in NQO1 expression regulation. Besides biotransformation, NQO1 also implicates in the maintenance of endogenous lipid-soluble antioxidants alpha-tocopherol and ubiquinone in their reduced/active forms, direct scavenging of superoxide anion radicals, stabilization of the tumor suppressor proteins including p53 and as a gatekeeper of the 20S proteasome pathway.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: NQO1