Anti-NQO1 Antibody Picoband™

Product Name

Anti-NQO1 Antibody Picoband™

View all NQO-1 Antibodies

SKU/Catalog Number

A00494-2

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-NQO1 Antibody Picoband™ catalog # A00494-2. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-NQO1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00494-2)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human NQO1 recombinant protein (Position: M1-K274).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A00494-2 is reactive to NQO1 in Human

Applications

A00494-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

31 kDa

Calculated molecular weight

29159 MW

Background of NQO-1

NAD(P)H dehydrogenase [quinone] 1 is an enzyme that in humans is encoded by the NQO1 gene. This gene is a member of the NAD(P)H dehydrogenase (quinone) family and encodes a cytoplasmic 2-electron reductase. And this FAD-binding protein forms homodimers and reduces quinones to hydroquinones. In addition, this protein's enzymatic activity prevents the one electron reduction of quinones that results in the production of radical species. Mutations in this gene have been associated with tardive dyskinesia (TD), an increased risk of hematotoxicity after exposure to benzene, and susceptibility to various forms of cancer. Altered expression of this protein has been seen in many tumors and is also associated with Alzheimer's disease (AD). Alternate transcriptional splice variants, encoding different isoforms, have been characterized.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

View more details

Reconstitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.25 µg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry (Fixed), 1-3 µg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

Click image to see more details

Figure 1. Western blot analysis of NQO1 using anti-NQO1 antibody (A00494-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human MCF-7 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human A549 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NQO1 antigen affinity purified polyclonal antibody (Catalog # A00494-2) at 0.25 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for NQO1 at approximately 31 kDa. The expected band size for NQO1 is at 31 kDa.

Click image to see more details

Figure 2. IF analysis of NQO1 using anti-NQO1 antibody (A00494-2).
NQO1 was detected in an immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-NQO1 Antibody (A00494-2) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Click image to see more details

Figure 3. Flow Cytometry analysis of MCF-7 cells using anti-NQO1 antibody (A00494-2).
Overlay histogram showing MCF-7 cells stained with A00494-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NQO1 Antibody (A00494-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Gene/Protein Information For NQO1 (Source: Uniprot.org, NCBI)

Alternative Names

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 NQO1 DHQU, DIA4, DTD, NMOR1, NMORI, QR1 NAD(P)H quinone dehydrogenase 1 NAD(P)H dehydrogenase [quinone] 1|DT-diaphorase|NAD(P)H dehydrogenase, quinone 1|NAD(P)H:Quinone acceptor oxidoreductase type 1|NAD(P)H:menadione oxidoreductase 1|NAD(P)H:quinone oxidoreductase 1|NAD(P)H:quinone oxireductase|azoreductase|diaphorase (NADH/NADPH) (cytochrome b-5 reductase)|diaphorase-4|dioxin-inducible 1|menadione reductase|phylloquinone reductase|quinone reductase 1

For more info on NQO1, check out the NQO1 Infographic

We have 30,000+ of these available, one for each gene! Check them out.

In this infographic, you will see the following information for NQO1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

Take me to the NQO1 Infographic

Here are featured tools and databases that you might find useful.

• Boster's Pathways Library
• Protein Databases
• Bioscience Research Protocol Resources
• Data Processing & Analysis Software
• Photo Editing Software
• Scientific Literature Resources
• Research Paper Management Tools
• Molecular Biology Software
• Primer Design Tools
• Bioinformatics Tools
• Phylogenetic Tree Analysis