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Synthetic peptide conjugated to KLH derived from within residues 200 to the C-terminus of Human NQO1.
(Peptide available as ab34172.)
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.
|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.|
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 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.
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