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. This polyclonal antibody has been reported for use in ELISA and ELISPOT.
Applications Tested
The biotinylated rabbit anti-mouse/rat TNF-α polyclonal antibody has been tested as the detection antibody in a sandwich ELISA for analysis of mouse or rat TNF-α in combination with the purified anti-mouse/rat TNF-α monoclonal antibody, clone TN3.19 (14-7423) for capture and recombinant mouse TNF-α (14-8321) or rat TNF-α (14-8321) as the standard. A suitable range of concentrations of this biotinylated antibody for ELISA detection is 0.5 – 2.0 µg/ml. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 2000 pg/ml - 15 pg/ml (mouse TNF-α) or 5000 – 30 pg/ml (rat TNF-α) should be included in each ELISA plate. Recognition of mouse TNF-α by this assay is not interfered with by 1000x excess soluble TNF-α receptor type I or II.
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.