Il1b Antibody

This is an IgG preparation of whole rabbit serum purified by DEAE fractionation. This antibody is primarily directed against mature, 17,000 MW mouse IL-1ß and is useful in determining its presence in various assays. The antibody does not recognize human IL-1ß or mouse IL-1α based on a neutralization assay. In ELISA formats and other immunoreactive assays, reactivity occurs with rat IL-1ß. This antibody will recognize 10% of the non-denatured (native) precursor 31,000 MW mouse IL-1ß containing samples but will primarily detect all of the 17,000 MW mature molecule. However, in immunoblot analysis, the usual procedure of heating the sample in SDS with or without reducing agents will facilitate denaturing of the 31,000 MW IL- 1ß precursor molecule. Denatured 31,000 precursor IL-1ß will be recognized by this antibody.

Store Anti-IL-1 beta antibody at 4° C prior to restoration. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.

Anti-Mouse IL-1ß has been tested for use in immunohistochemistry, immunoblotting and immunofluorescence. This antibody is useful in ELISA, neutralizations, radioimmunoassays, flow cytometry, and immunoprecipitation. It recognizes the 17,000 MW mature IL-1ß. For immunoblots, typically, IL-1ß is detected from supernatants or lysates of 2 x 10E6 endotoxin-stimulated peripheral blood mononuclear cells (PBMC) . PBMC are stimulated for 24 hours with 1% (v/v) serum plus 10 ng/mL E.coli LPS. For immunoprecipitation pre-clearing the preparation with a non-specific Rabbit IgG to reduce background is suggested. For immunohistochemistry either paraffin fixation or cryofixation can be used for sample preparation to stain intracellular IL-1ß. For ELISA use HRP Conjugated Anti-Rabbit IgG [H&L] (Goat) for detection. In ELISA formats this antibody is best used as the second antibody in combination with a monoclonal antibody as a capture antibody. This antibody is also useful for neutralization of mouse and rat IL-1ß activity in bioassays. It does not neutralize the biological activity IL-1α. It does not neutralize the biological activity of human or primate IL-1ß. For neutralization, it is recommended to incubate the sample with a dilution of the antibody for at least 4 hours before being tested. A control of similarly diluted normal rabbit IgG is recommended. This antibody can be used for FACS analysis. Caution should be exhibited as the F (c) domain of the rabbit IgG molecule may interact with cells non-specifically. Reconstitution Buffer: Restore with deionized water (or equivalent) . Reconstitution Volume: 100 μL