Determination of IL12

The determination of IL-12 can be carried out by biological assays or by ELISA. Biological assays for IL-12 are very sensitive and are mostly based on the 'capture' technique. The assay wells are precoated with an antibody against IL-12 that should be specific for the heter-odimer, but not neutralizing and not binding to IL-12p40 or IL-12(p40)2. After capturing IL-12 the responding cells (e.g. activated T cells) are added and the resulting IFNy production or proliferation can be measured. This avoids, especially in the mouse system, inhibitory effects of IL-12(p40)2, which may be also present in the test fluids. A sandwich ELISA uses the same primary antibody to bind IL-12 but not the p40 chain that is usually produced in excess over IL-12. A

secondary anti-IL-12p40 or anti-IL-12p35 antibody can then be used for detection of the IL-12 bound.

See also: Adjuvants; Autoimmunity; CD40 and its ligand; Helper T lymphocytes; Cytokine receptors; Dendritic cells; Immunotherapy of tumors; Innate immunity; Interferon y receptor; Interleukin 6 receptor; Macrophage activation; Natural killer (NK) cells; Septic shock.

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