Product datasheet

Anti-NQO1 antibody ab34173

9 Abreviews 57 References 6 Images

Overview

Product name Anti-NQO1 antibody

Description Rabbit polyclonal to NQO1
Host species Rabbit
Tested applications Suitable for: Sandwich ELISA, IHC-P, IHC-Fr, WB, ELISA, ICC/IF
Species reactivity Reacts with: Mouse, Rat, Guinea pig, Human
Immunogen Synthetic peptide conjugated to KLH derived from within residues 200 to the C-terminus of
Human NQO1. Read Abcam's proprietary immunogen policy (Peptide available as ab34172.)

Positive control Recombinant Human NQO1 protein (ab87692) can be used as a positive control in WB. This
antibody gave a positive signal in the following whole tissue lysates: Human Kidney Normal. This
antibody gave a positive signal in the following tissues: Formalin Fixed Paraffin Embedded
Human Breast Cancer.

Properties

Form Liquid
Storage instructions Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -
80°C. Avoid freeze / thaw cycle.
Storage buffer Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS. pH 7.4
Purity Immunogen affinity purified
Clonality Polyclonal
Isotype IgG

Applications

Our Abpromise guarantee covers the use of ab34173 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

1
Application Abreviews Notes

Sandwich 1/1000.
ELISA

IHC-P 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

IHC-Fr Use at an assay dependent concentration.

WB Use a concentration of 1 µg/ml. Detects a band of approximately 31 kDa (predicted molecular weight: 31
kDa).Can be blocked with Human NQO1 peptide (ab34172).

ELISA Use at an assay dependent concentration. Can be paired for ELISA with Mouse monoclonal [A180] to
NQO1 (ab28947).

ICC/IF Use a concentration of 1 µg/ml.

Target

Function
Sequence similarities
Cellular localization
The enzyme apparently serves as a quinone reductase in connection with conjugation reactions of
hydroquinons involved in detoxification pathways as well as in biosynthetic processes such as the
vitamin K-dependent gamma-carboxylation of glutamate residues in prothrombin synthesis.
Belongs to the NAD(P)H dehydrogenase (quinone) family.
Cytoplasm.

Images

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)

Lane 2: NQO1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HepG2 whole cell lysate (20 µg)
Lane 4: MCF7 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab34173
observed at 31 kDa. Red - loading control, ab18058, observed at
130 kDa.

ab34173 was shown to specifically react with NQO1 in wild-type

HAP1 cells as signal was lost in NQO1 knockout cells. Wild-type
and NQO1 knockout samples were subjected to SDS-PAGE.
ab34173 and ab18058 (Mouse anti-Vinculin loading control) were
Western blot - Anti-NQO1 antibody (ab34173)
incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution
respectively. Blots were developed with Goat anti-Rabbit IgG H&L
(IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG
H&L (IRDye® 680RD) preabsorbed ab216776 secondary
antibodies at 1/20000 dilution for 1 hour at room temperature
before imaging.

2
 Anti-NQO1 antibody (ab34173) at 1 µg/ml + Human Kidney Whole
Tissue Lysate at 20 µg

Secondary
IRDye 680 Conjugated Goat Anti Rabbit IgG (H&L) at 1/15000
dilution

Performed under reducing conditions.

Predicted band size: 31 kDa

Observed band size: 31 kDa
Western blot - Anti-NQO1 antibody (ab34173)

Image courtesy of Human Protein Atlas . ab34173 staining NQO1 in

human breast cancer. Paraffin embedded human breast cancer
tissue was incubated with ab34173 (1/200 dilution) for 30 mins at
room temperature. Antigen retrieval was performed by heat
induction in citrate buffer pH 6.

ab34173 was tested in a tissue microarray (TMA) containing a wide

range of normal and cancer tissues. Further results for this antibody
can be found at www.proteinatlas.org

Immunohistochemistry (Formalin/PFA-fixed paraffin-

embedded sections) - Anti-NQO1 antibody
(ab34173)

ICC/IF image of ab34173 stained HeLa cells. The cells were 4%

formaldehdye fixed (10 min) and then incubated in 1%BSA / 10%
normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to
permeabilise the cells and block non-specific protein-protein
interactions. The cells were then incubated with the antibody
(ab34173, 1µg/ml) overnight at +4°C. The secondary antibody
(green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L)
used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to
label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was
used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Immunocytochemistry/ Immunofluorescence - Anti-

NQO1 antibody (ab34173)

3
 Sandwich ELISA for the detection of NQO1, using ab28947 (1/500)
as the cature antibody and ab34173 (1/1000) for the detection.
Please refer to abreview for further experimental details.

Sandwich ELISA - Anti-NQO1 antibody (ab34173)

This image is courtesy of an anonymous Abreview

Anti-NQO1 antibody (ab34173) at 1 µg/ml + Recombinant human

NQO1 protein (ab59663) at 0.01 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at
1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Western blot - Anti-NQO1 antibody (ab34173) Predicted band size: 31 kDa

Exposure time: 30 seconds

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