IL1RA (IL1RN) Rabbit Polyclonal Antibody

Specifications

Product Data
Applications	ELISA, FN, IHC, WB
Recommend Dilution	This IgG fraction antibody of anti-Human IL-1 Receptor Antagonist has been tested for use in Neutralizations, ELISA, Radioimmunoassays, Western blotting, Immunohistochemistry and Flow cytometry. Reactivity in other immunoassays is unknown. Note: This product has been assayed by immunoblot using HRP Goat-anti-Rabbit IgG [H&L] and TMB as a substrate. A working dilution range of 1:400 to 1:800 is suggested for this application. This product has been assayed by ELISA against IL-1Receptor Antagonist using HRP Conjugated Anti-Rabbit IgG [H&L] (Goat) and ABTS as a substrate for 30 minutes at room temperature. A working dilution range of 1:500 to 1:1,000 is suggested for this product. For use in ELISA formats, this antibody is best used as the second antibody in combination with a monoclonal antibody as a capture antibody. This product has been assayed by Immunohistochemistry. A dilution of 1:400 or greater is suggested for this immunoassay. Either paraffin fixation or cryofixation can be used for Immunohistochemistry. This product has been assayed by radioimmunoassay against antigen. A dilution of 1:8,000 is suggested for this immunoassay. Recommended Dilutions: ELISA: 1/1,000-1/5,000. Western Blot: 1/500-1/2,000.
Reactivity	Human, Monkey
Host	Rabbit
Clonality	Polyclonal
Immunogen	Prepared by repeated immunizations with recombinant human sIL-1Ra produced in E.coli.
Specificity	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 10e6 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. In ELISA formats and other immunoreactive assays, this antibody will recognize both sIL-1Ra and icIL-1Ra. Biologically, sIL-1Ra and icIL-1Ra both function as IL-1 receptor antagonists by binding to the type I IL-1R and prevent signaling by IL-1a or IL-1b . However, sIL-1Ra is primarily secreted whereas icIL-1Ra remains associated with the cell. This antibody is also useful for neutralization of human and primate IL-1Ra. If a body fluid, cell supernatant or serum contains IL-1Ra, it is possible to neutralize the inhibitory activity of IL-1Ra on IL-1 activity, being exogenous or endogenous IL-1. A 1:200 dilution of this antibody is incubated with the sample for 4 hours. The mixture is then added to cells in the assay for IL-1 activity. A control of normal rabbit IgG is recommended. An increase in the activity of the sample indicates that the inhibitory activity of IL-1Ra on an IL-1-mediated read-out has been neutralized. Using recombinant human IL-1Ra as a positive control, a 1:200 dilution of this antibody will neutralize the inhibitory activity of IL-1Ra at 100 ng/mL. This antibody does not neutralize the biological activity IL-1. 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.
Formulation	0.02M Potassium Phosphate, 0.15M Sodium Chloride, pH 7.2 with 0.01% (w/v) Sodium Azide as preservative. State: Purified State: Liquid (sterile filtered) purified Ig fraction.
Concentration	lot specific
Purification	DEAE Chromatography.
Conjugation	Unconjugated
Storage Condition	Store the antibody (undiluted) at 2-8°C for one month or (in aliquots) at -20°C for longer. Dilute only prior to immediate use. Avoid cycles of freezing and thawing.
Gene Name	interleukin 1 receptor antagonist
Database Link	Entrez Gene 3557 Human UniProt ID P18510
Background	IL1RA is a member of the Interleukin 1 cytokine family. This protein inhibits the activities of Interleukin 1, alpha (IL1A) and Interleukin 1, beta (IL1B), and modulates a variety of Interleukin 1 related immune and inflammatory responses. This gene and five other closely related cytokine genes form a gene cluster spanning approximately 400 kb on chromosome 2. A polymorphism of this gene is reported to be associated with increased risk of osteoporotic fractures and gastric cancer. Four alternatively spliced transcript variants encoding distinct isoforms have been reported.
Synonyms	IL1F3, IL-1ra, IL-1RN, IRAP, IL1 inhibitor, ICIL-1RA
Note	Endotoxin Content: < 10 pg/ml by LAL method.
Reference Data

*Delivery time may vary from web posted schedule. Occasional delays may occur due to unforeseen complexities in the preparation of your product. International customers may expect an additional 1-2 weeks in shipping.