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Recombinant full length protein of Human NQO1.
Alternative versions available:
Our Abpromise guarantee covers the use of ab28947 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||1/500. Can be used as capture antibody in conjunction with ab34173 as detection antibody.|
|IP||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 30 kDa.|
|ICC/IF||Use a concentration of 1 µg/ml.|
|ELISA||Use at an assay dependent concentration. Can be paired for ELISA with Rabbit polyclonal to NQO1 (ab34173).|
|IHC-P||Use at an assay dependent concentration.|
IHC image of NQO1 staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab28947, 0.1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ICC/IF image of ab28947 stained human HEK 293 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab28947, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Overlay histogram showing HeLa cells stained with ab28947 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab28947, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was goat anti-mouse DyLight® 488 (IgG H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Immunohistochemical analysis of dog skin tissue, staining NQO1 with ab28947.
Tissue was fixed with formaldehyde and antigen retrieval was by heat mediation in a citrate buffer (pH 6). Samples were incubated with primary antibody (1/175 in BSA in TBS) for 45 minutes. ab98784 rabbit polyclonal to anti-mouse HRP (IgG (1/500) was used as the secondary antibody.
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