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Our Abpromise guarantee covers the use of ab34674 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||1/400 - 1/800.|
|WB||1/400 - 1/800. Detects a band of approximately 17 kDa (predicted molecular weight: 26 kDa).Can be blocked with Synthetic Mouse TNF alpha protein (ab151311). Detects TNF alpha from supernatants or lysates of 2 x 106 endotoxin stimulated mouse peripheral blood mononuclear cells (PBMC).|
|IHC-P||Use at an assay dependent concentration.|
|ELISA||1/1000 - 1/2000. For use in ELISA formats, this antibody is best used as the second antibody in combination with a monoclonal antibody as a capture antibody.|
|Neutralising||1/200. It is recommended to incubate the sample with the dilution of the antibody for at least 4 hours before being tested. A control of similarly diluted normal rabbit IgG is recommended.|
Immunohistochemical analysis of frozen PFA perfusion-fixed murine dermal tissue sections, labelling TNF with ab34674 at a concentration of 1/200 incubated for 24 hours at 4°C in TBS. Permeablization was with Triton X-100. Blocking was with 5% serum incubated for 30 minutes at 22°C. The secondary was a polyclonal Donkey anti-rabbit Cy3® conjugate at 1/200.
ab34674 staining TNF alpha in mouse knee joint sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed. Samples were then blocked with 10 minutes at room temperature followed by incubation with the primary antibody at a 1/200 dilution for 30 minutes. An undiluted HRP-conjugated human anti-rabbit monoclonal was used as secondary antibody.