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Recombinant fragment corresponding to Human TNF alpha aa 77-233. E.coli derived Recombinant Human TNF-alpha
Database link: P01375
Our Abpromise guarantee covers the use of ab9635 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|WB||Use at an assay dependent concentration. To detect TNF-alpha by Western Blot analysis this antibody can be used at a concentration of 0.1 - 0.2 µg/ml. Used in conjunction with compatible secondary reagents the detection limit for recombinant TNF-alpha is 1.5 - 3.0 ng/lane, under either reducing or non-reducing conditions.|
|ELISA||Use at an assay dependent concentration. Can be paired for ELISA with Mouse monoclonal to TNF alpha (ab9348). To detect TNF-alpha by direct ELISA (using 100µl/well antibody solution) a concentration of at least 0.5µg/ml of this antibody is required. This antigen affinity purified antibody, in conjunction with compatible secondary reagents, allows the detection of 0.2 - 0.4 ng/well of recombinant TNF-alpha.|
|Neutralising||Use at an assay dependent concentration. To yield one-half maximal inhibition [ND50] of the biological activity of hTNF-alpha (0.5 ng/ml), a concentration of 0.08 - 0.10 µg/ml of this antibody is required.|
|Sandwich ELISA||Use a concentration of 0.5 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal [2C8] to TNF alpha (ab8348). For sandwich ELISA, use this antibody as Detection at 0.5µg/ml with Ab8348 as Capture.|
IHC image of TNF alpha staining in human tonsil 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 ab9635, 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.