IL-6 antibody has been tested for use in neutralizations, ELISA, immunohistochemistry, radioimmunoassay, immunoprecipitation, and immunoblot. Reactivity in other immunoassays is unknown. Expect ~23.7kDa. In Western blot analysis of natural cell products or human body fluids, multiple bands of IL-6 will appear due to the variable amount of glycosylation on the molecule. For immunoblot use the supernatants or lysates of 2 x 106 endotoxin-stimulated human peripheral blood mononuclear cells (PBMC). PBMC are stimulated for 24 hours with 1% (v/v) human serum plus 10ng/mL E. coli LPS. For immunoprecipitation, pre-clearing with a non-specific rabbit IgG is helpful to reduce background. The antiserum is useful for neutralization of human of IL-6 activity in bioassays. For neutralization, incubate the sample with a 1:400 dilution of the antiserum for at least 4 hours before being tested. A control of similarly diluted normal rabbit IgG (heat inactivated) is recommended. In neutralization experiments in vitro, this antibody does not result in enhanced activity of IL-6. However, because antibodies to IL-6 may act as a soluble receptor in vivo, some antibodies to IL-6 act as carriers and enhance IL-6 activity. For immunohistochemistry either paraffin fix or cryofix tissue.