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NQO1 Recombinant Rabbit Monoclonal Antibody [JF440-1] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750349

NQO1 Recombinant Rabbit Monoclonal Antibody [JF440-1] - BSA and Azide free

Application

  • WB

  • IP

  • FC

  • IHC-P

Reactivity

  • Human

  • Mouse

With BSA and Azide
Predicted reactivity

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

NQO1 Recombinant Rabbit Monoclonal Antibody [JF440-1] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human NQO1 aa 1-49 / 274.

Species Reactivity

Human, Mouse (Predicted: Rat)

Predicted species support after-sales service

Validated Applications

WB, IP, FC, IHC-P

Target Molecular Weight

Predicted band size: 31 kDa

Positive Control

HCT 116 cell lysate, HepG2 cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, Mouse kidney tissue lysate, Rat kidney tissue lysate, human breast cancer tissue, MCF7.

Conjugation

unconjugated

Clone Number

JF440-1

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • IP

  • Use at an assay dependent concentration.

  • FC

  • 1:1,000

  • IHC-P

  • 1:1,000

Target

Function

NAD(P)H:quinone oxidoreductase 1 (NQO1) and NRH:quinone oxidoreductase (NQO2) are flavoproteins that catalyze the metabolic detoxification of quinones and their derivatives to hydroquinones, using either NADH or NADPH as the electron donor. This protects cells against quinone-induced oxidative stress, cytotoxicity, and mutagenicity. Many tumors overexpress NQO1, which is an obligate two-electron reductase that deactivates toxins and activates bioreductive anticancer drugs. NQO1, a 274 amino acid protein, is ubiquitously expressed, but the expression level varies among tissues. NQO1 gene expression is coordinately induced in response to xenobiotics, antioxidants, heavy metals and radiation. The antioxidant response element (ARE) in the NQO1 gene promoter is essential for expression and coordinated induction of NQO1. ARE activation by tert-butylhydroquinone is dependent on PI3-kinase, which lies upstream of Nrf2. Nrf2, c-Jun, Nrf1, Jun-B and Jun-D bind to the ARE and regulate expression and induction of NQO1 gene. Maf-Maf homodimers and possibly Maf-Nrf2 heterodimers play a role in negative regulation of ARE-mediated transcription, but Maf-Nrf1 heterodimers fail to bind with the NQO1 gene ARE and do not repress NQO1 transcription.

Background References

1. Greco T et al. Ketogenic diet decreases oxidative stress and improves mitochondrial respiratory complex activity. J Cereb Blood Flow Metab 36:1603-13 (2016).

2. Kigoshi Y et al. CACUL1/CAC1 Regulates the Antioxidant Response by Stabilizing Nrf2. Sci Rep 5:12857 (2015).

Sequence Similarity

Belongs to the NAD(P)H dehydrogenase (quinone) family.

Subcellular Location

Cytoplasm.

UNIPROT

SwissProt: P15559 Human

SwissProt: Q64669 Mouse

SwissProt: P05982 Rat

Synonyms

Azoreductase antibody

Cytochrome b 5 reductase antibody

DHQU antibody

DIA 4 antibody

DIA4 antibody

Diaphorase (NADH/NADPH) (cytochrome b 5 reductase) antibody

Diaphorase (NADH/NADPH) antibody

Diaphorase 4 antibody

Dioxin inducible 1 antibody

DT diaphorase antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of NQO1 on different lysates with Rabbit anti-NQO1 antibody (<a href="/products/HA750349" style="font-weight: bold;text-decoration: underline;">HA750349</a>) at 1/2,000 dilution.<br /><br />Lane 1: HCT 116-si NT cell lysate<br />Lane 2: HCT 116-si NQO1 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 31 kDa<br />Observed band size: 31 kDa<br /><br />Exposure time: 16 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750349" style="font-weight: bold;text-decoration: underline;">HA750349</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of NQO1 on different lysates with Rabbit anti-NQO1 antibody (HA750349) at 1/2,000 dilution.

    Lane 1: HCT 116-si NT cell lysate
    Lane 2: HCT 116-si NQO1 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 31 kDa
    Observed band size: 31 kDa

    Exposure time: 16 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750349) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of NQO1 on different lysates with Rabbit anti-NQO1 antibody (<a href="/products/HA750349" style="font-weight: bold;text-decoration: underline;">HA750349</a>) at 1/1,000 dilution.<br /><br />Lane 1: HepG2 cell lysate (20 µg/Lane)<br />Lane 2: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 3: C2C12 cell lysate (20 µg/Lane)<br />Lane 4: Mouse kidney tissue lysate (40 µg/Lane)<br />Lane 5: Rat kidney tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 31 kDa<br />Observed band size: 31 kDa<br /><br />Exposure time: 4 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750349" style="font-weight: bold;text-decoration: underline;">HA750349</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of NQO1 on different lysates with Rabbit anti-NQO1 antibody (HA750349) at 1/1,000 dilution.

    Lane 1: HepG2 cell lysate (20 µg/Lane)
    Lane 2: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 3: C2C12 cell lysate (20 µg/Lane)
    Lane 4: Mouse kidney tissue lysate (40 µg/Lane)
    Lane 5: Rat kidney tissue lysate (40 µg/Lane)

    Predicted band size: 31 kDa
    Observed band size: 31 kDa

    Exposure time: 4 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750349) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-NQO1 antibody (<a href="/products/HA750349" style="font-weight: bold;text-decoration: underline;">HA750349</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750349" style="font-weight: bold;text-decoration: underline;">HA750349</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-NQO1 antibody (HA750349) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750349) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of MCF7 cells labeling NQO1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA750349" style="font-weight: bold;text-decoration: underline;">HA750349</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of MCF7 cells labeling NQO1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA750349, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"