Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
156-10000 pg/mL
Sample Type
Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Mouse IL-1ra/IL-1F3 DuoSet ELISA Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Background: IL-1ra/IL-1F3
Mouse Interleukin-1 receptor antagonist (IL-1ra) is a 22-25 kDa glycoprotein produced by a
variety of cell types that antagonizes IL-1 activity (1-3). It is a member of the IL-1 family of
proteins that includes IL-1 alpha and IL-1 beta. Although there is little amino acid (aa) identity (< 30%)
among the three IL-1 family members, all molecules bind to the same receptors, all show a
beta -trefoil structure, all are physically placed on mouse chromosome # 2, and all are believed to
have evolved from a common ancestral gene (1-4). Mouse IL-1ra is synthesized as a 178 aa
precursor that contains a 26 aa signal sequence plus a 152 aa mature region. There is one
intrachain disulfide bond and one potential N-linked glycosylation site (3, 5, 6). Mature mouse
IL-1ra shares 90%, 77%, 80%, 80%, and 78% aa sequence identity to mature rat (4), human (7),
porcine (8), canine (9), and equine (10) IL-1ra, respectively. In humans, three non-secreted IL-1ra
isoforms have also been identified (11-14). These result from the use of alternate start sites or
exon splicing. In mice, only one of the three human intracellular isoforms has been isolated.
This mouse molecule is the ortholog of the 159 aa human intracellular isoform # 1 (6, 12). The
mouse intracellular form differs from the secreted precursor by only three amino acids. Cells
known to secrete IL-1ra include dermal fibroblasts (15), vascular smooth muscle cells (16),
intestinal columnar epithelium (17), chondrocytes (18), macrophages (19), non-keratinized oral
stratified squamous epithelium (20), mast cells (21), neutrophils and monocytes (22), Sertoli
cells (23), and hepatocytes (24). There are two type I transmembrane glycoprotein receptors for IL-1ra: the bioactive 80 kDa
type I IL-1 receptor (IL-1 RI), and the inert (decoy) 65 kDa type II IL-1 receptor (IL-1 RII). IL-1ra
binding to IL-1 RI competitively blocks IL-1 ( alpha or beta ) binding to the same receptor. Unlike the
IL-1/IL-1 RI complex, the IL-1ra/IL-1 RI complex cannot recruit the IL-1 receptor accessory
protein that is required for signal transduction. This results in receptor ligation without cell
activation (1, 25). IL-1ra also competitively blocks IL-1 binding to the decoy IL-1 RII. In this case,
IL-1ra may actually potentiate IL-1 activity by cancelling the functions of both antagonists.
All activities attributed to IL-1ra are explained by its role as a competitive inhibitor of IL-1
binding to IL-1 RI (1, 2, 26, 27). In general, the ratio of IL-1ra to IL-1 beta is close to 1 in both health
and disease (26, 28). To achieve total abrogation of IL-1 activity, more than 95% of all IL-1 RI
receptors apparently need to be occupied by IL-1ra (29). Thus, local concentrations of secreted
IL-1ra may be the critical determinants of IL-1 activity. The function of intracellular IL-1ra is less
clear. While it would seem to be a simple competitor of IL-1 activity, its role may be limited to
that of downmodulating non-specific inflammation associated with cell debris and death (14,
27). It has no action intracellularly; only when released through cell death would it then
become a functional IL-1 antagonist.
Long Name
Interleukin 1 Receptor Antagonist
Alternate Names
DIRA, ICIL-1ra, IL-1F3, IL-1ra3, IL-1RN, IL1ra, IL1RN, MVCD4
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