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NQO1 Recombinant Rabbit Monoclonal Antibody [JF440-1]

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Catalog# ET1702-50

NQO1 Recombinant Rabbit Monoclonal Antibody [JF440-1]

Application

  • WB

  • IP

  • FC

  • IHC-P

Reactivity

  • Human

  • Mouse

BSA and Azide free
Predicted reactivity

  • Rat

预测的种属支持售后
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

概述

产品名称

NQO1 Recombinant Rabbit Monoclonal Antibody [JF440-1]

抗体类型

Recombinant Rabbit monoclonal Antibody

免疫原

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

种属反应性

Human, Mouse (Predicted: Rat)

预测的种属支持售后

验证应用

WB, IP, FC, IHC-P

分子量

Predicted band size: 31 kDa

阳性对照

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.

偶联

unconjugated

克隆号

JF440-1

RRID

AB_3070312

产品特性

形态

Liquid

浓度

存放说明

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

存储缓冲液

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

亚型

IgG

纯化方式

Protein A affinity purified.

应用稀释度

  • WB

  • 1:1,000-1:5,000

  • IP

  • Use at an assay dependent concentration.

  • FC

  • 1:1,000

  • IHC-P

  • 1:1,000

靶点

功能

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.

背景文献

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).

序列相似性

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

亚细胞定位

Cytoplasm.

UNIPROT

SwissProt: P15559 Human

SwissProt: Q64669 Mouse

SwissProt: P05982 Rat

别名

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

展开

图片

  • <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/ET1702-50" style="font-weight: bold;text-decoration: underline;">ET1702-50</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/ET1702-50" style="font-weight: bold;text-decoration: underline;">ET1702-50</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 (ET1702-50) 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 (ET1702-50) 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/ET1702-50" style="font-weight: bold;text-decoration: underline;">ET1702-50</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/ET1702-50" style="font-weight: bold;text-decoration: underline;">ET1702-50</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 (ET1702-50) 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 (ET1702-50) 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/ET1702-50" style="font-weight: bold;text-decoration: underline;">ET1702-50</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/ET1702-50" style="font-weight: bold;text-decoration: underline;">ET1702-50</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 (ET1702-50) 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 (ET1702-50) 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/ET1702-50" style="font-weight: bold;text-decoration: underline;">ET1702-50</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 (ET1702-50, 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).

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

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