ELISA, Immunohistochemistry, Immunoprecipitation, Western Blot. IL-6 antibody has been tested for use in neutralizations, ELISA, immunohistochemistry, radioimmunoassay, immunoprecipitation, and immunoblotting. Reactivity in other immunoassays is unknown. For immunoblotting 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 10 ng/mL E.coli LPS. For immunohistochemistry either paraffin fix or cryofix tissue. For immunoprecipitation, pre-clearing with a non-specific rabbit IgG is helpful to reduce background.
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Description
Anti-IL-6 antiserum detects recombinant and native IL-6 present in body fluids and cell supernatants in various assays (ie. IL-1 stimulated IL-6 production from fibroblasts). 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. The antiserum is also 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. This product has minimal reactivity with mouse IL-6.
Immunogen
This whole rabbit serum was prepared by repeated immunizations with recombinant human IL-6 produced in E.coli.