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Our Abpromise guarantee covers the use of ab76997 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 56 kDa.|
|ELISA||Use at an assay dependent concentration. Detection limit for ab76997 is approximately 30 ng/ml when used as a capture antibody.|
|IHC-P||Use a concentration of 0.7 µg/ml. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.|
|ICC/IF||Use a concentration of 10 µg/ml.|
|Flow Cyt||Use 1µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|IP||Use at an assay dependent concentration.|
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling SOX9 with ab76997 at 0.7 µg/ml.
Immunocytochemistry/Immunofluorescence analysis of SOX9 expression in HepG2 cells, using 10 µg/ml of ab76997
Overlay histogram showing HepG2 cells stained with ab76997 (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 (ab76997, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Immunoprecipitation of SOX9 transfected lysate using anti-SOX9 monoclonal antibody and Protein A magnetic beads. Western blot was performed with an rabbit anti-SOX9 polyclonal antibody.