Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µL to 5 µL. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
Description
The affinity purified polyclonal rabbit antibody reacts with mouse and rat tumor necrosis factor-alpha. Mouse TNF-α is a 17 kDa factor produced by macrophages, monocytes, neutrophils, CD4 T cells, and NK cells. A 26 kDa form is expressed as a membrane-bound molecule. TNF-α is cytolytic and plays an important role in immune regulation. Bioactive TNF-α exists as a homotrimer.
Applications Reported
For research use only, not for diagnostic or therapeutic use. The Affinity Purified anti-mouse/rat TNF-α anti-mouse TNF-a antibody has been reported for use as the plate-coating (ELISA Capture) antibody for the In Vivo Cytokine Capture Assay (IVCCA), in conjunction with the Functional Grade Biotinylated TN3.19 antibody for In Vivo Capture (ELISA Detection). The IVCCA facilitates measurement of cytokines in serum by increasing their in vivo half lives dramatically. This increases the sensitivity of measurement of in vivo cytokine production 30- to 1000- fold.
For detailed methodology in this application, please refer to this reference: Finkelman, F., S. Morris, T. Orekhova, and D. Sehy. 2003. The In Vivo Cytokine Capture Assay for measurement of cytokine production in the mouse. In Current Protocols in Immunology. Unit 6.28. J. Coligan, A. Kruisbeek, D. Margulies, E. Shevach, and W. Strober, eds. John Wiley and Sons, New York.
Applications Tested
The Affinity Purified anti-mouse/rat TNF-α antibody has been tested as the plate coating ELISA capture antibody in a sandwich ELISA for analysis of mouse TNF-a, in combination with the Functional Grade Biotinylated anti-mouse TNF-a antibody (14-8321) for ELISA detection and recombinant mouse TNF-a (14-8321) as the standard. A suitable range of concentrations of the Affinity Purified anti-mouse/rat TNF-α antibody for ELISA capture is 1.0 - 4.0 µg/ml. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 2000 pg/ml - 15.0 pg/ml should be included in each ELISA plate.
References
Finkelman, F., S. Morris, T. Orekhova, and D. Sehy. 2003. The In Vivo Cytokine Capture Assay for measurement of cytokine production in the mouse. In Current Protocols in Immunology. Unit 6.28. J. Coligan, A. Kruisbeek, D. Margulies, E. Shevach, and W. Strober, eds. John Wiley and Sons, New York.
Finkelman, F.D., and S.C. Morris. 1999. Development of an assay to measure in vivo cytokine production in the mouse. Int. Immunology. 11: 1811-1818.