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Our Abpromise guarantee covers the use of ab9732 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 0.1 - 0.2 µg/ml.|
|ELISA||Use a concentration of 0.5 µg/ml. Allows the detection of 0.2 - 0.4 ng/well of recombinant mIL-7 (using 100 µl/well antibody solution).|
|Neutralising||Use a concentration of 0.08 - 0.1 µg/ml. for one-half maximal inhibition [ND50] of the biological activity of mIL-7 (0.50 ng/ml).|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
IHC image of IL7 staining in mouse spleen formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol B. 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 ab9732, 5µg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC 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.