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Mouse IL-1ra/IL-1F3 DuoSet ELISA

Bio-Techne includes R&D Systems | Catalog # DY480

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DY480

Ancillary Products Available

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

Reactivity

Mouse

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

Product Summary for Mouse IL-1ra/IL-1F3 DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse IL-1ra/IL-1F3. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Specifications

Assay Format

96-well strip plate (sold separately)

Sample Volume Required

100 µL

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Label

HRP

Scientific Data Images for Mouse IL-1ra/IL-1F3 DuoSet ELISA

Mouse IL-1ra / IL-1F3 ELISA Standard Curve

Mouse IL-1ra / IL-1F3 ELISA Standard Curve

Kit Contents for Mouse IL-1ra/IL-1F3 DuoSet ELISA

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

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)

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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

Entrez Gene IDs

3557 (Human); 16181 (Mouse); 60582 (Rat); 397499 (Porcine)

Gene Symbol

IL1RN

Additional IL-1ra/IL-1F3 Products

Product Documents for Mouse IL-1ra/IL-1F3 DuoSet ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse IL-1ra/IL-1F3 DuoSet ELISA

For research use only

Citations for Mouse IL-1ra/IL-1F3 DuoSet ELISA (19)

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  • Mouse IL-1ra/IL-1F3 DuoSet ELISA
    Name: Anonymous
    Sample Tested: peritoneal lavage fluid
    Verified Customer | Posted 04/15/2019
    Mice (HIII and LIII strains) injected i.p. with 2,4,6,10 tetramethylpentadecane (pristane).
    Mouse IL-1ra/IL-1F3 DuoSet ELISA DY480

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Protocols

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