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Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab185966 as a replacement.
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.
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