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Our Abpromise guarantee covers the use of ab28538 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 10 µg/ml.|
|IHC-P||Use at an assay dependent concentration.|
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 38 kDa.|
|Flow Cyt||Use 1µg for 106 cells. ab18407-Rat monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
ab28538 staining PAX9 in mouse E11.5 embryo tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with 2% paraformaldehyde in PBS for 2 hours at 4°C then placed in 30% sucrose overnight at 4°C prior to embedding in OCT using liquid nitrogen. Tissue was blocked with 1X PBS + 1.5% Milk + 1.5% BSA + 0.1% Triton X-100 for 12 hours at 4°C. Samples were incubated with primary antibody (1/500 in blocking buffer) for 1 hour at 23°C. An Alexa Fluor® 568-conjugated goat anti-rat IgG polyclonal (1/250) was used as the secondary antibody.
Overlay histogram showing HepG2 cells stained with ab28538 (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 (ab28538, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was goat anti-rat DyLight® 488 (IgG H+L) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG1, kappa monoclonal [RTK2071] (ab18412, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.