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Background
Interleukin 1 receptor antagonist is secreted by various types of cells including immune, epithelial and adipocytes. IL 1Ra is used to treat rheumatoid arthritis. Anti-IL-1 RA antibody is ideal for investigators involved in Cancer and Immunology research.
The whole rabbit serum used to produce this IgG fraction antibody was prepared by repeated immunizations with recombinant human sIL-1Ra produced in E.coli.
This is an IgG preparation of whole rabbit serum purified by DEAE chromatography. In general, this antibody also detects primate IL-1Ra in the same formats using similar dilutions, but not mouse or rabbit IL-1Ra. The antibody does not recognize human IL-1a, IL-1b or IL-1 receptor type I and type II. This antibody is primarily directed against the secreted form of IL-1Ra (also termed sIL-1Ra) but recognizes the native (non-denatured) as well as denatured intracellular form of IL-1Ra (also termed icIL-1Ra). IcIL-1Ra is found in many cells as constitutively expressed protein, particularly epithelial cells and keratinocytes. This antibody does not distinguish between sIL-1Ra and icIL-1Ra in ELISA formats or by immunohistochemistry. When used in immunoblot analysis, it will recognize 17,000 MW IL-1Ra in supernatants released from 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. However, in lysates of PBMC, icIL-1Ra is also found after 48-72 hours of culture. In monocytic cells, the initial IL-1Ra transcript is for sIL-1Ra which can be found almost exclusively intracellularly.
This IgG fraction antibody of anti-Human IL-1 Receptor Antagonist has been tested in western blot and suitable for use in neutralizations, ELISA, radioimmunoassays, immunohistochemistry and flow cytometry. Reactivity in other immunoassays is unknown. This product has been assayed by immunoblot using HRP Goat-anti-Rabbit IgG [H&L] (code # 611-1302) and TMB as a substrate. A working dilution range of 1:400 to 1:800 is suggested for this application. A working dilution range of 1:500 to 1:1,000 is suggested for ELISA. For use in ELISA formats, this antibody is best used as the second antibody in combination with a monoclonal antibody as a capture antibody. A dilution of 1:400 or greater is suggested for immunohistochemistry. Either paraffin fixation or cryofixation can be used for immunohistochemistry. A dilution of 1:8,000 is suggested for radioimmunoassay against antigen.
Formulation
Liquid (sterile filtered)
1.0 mg/mL by UV absorbance at 280 nm
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
0.01% (w/v) Sodium Azide
None
Shipping & Handling
Dry Ice
Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. 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.
Expiration date is one (1) year from date of receipt.
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
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