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Mouse IL-1ra/IL-1F3 Quantikine ELISA Kit

Bio-Techne includes R&D Systems | Catalog # MRA00

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MRA00

Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)

Sensitivity

13 pg/mL

Assay Range

31.2-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)

Product Summary for Mouse IL-1ra/IL-1F3 Quantikine ELISA Kit

The Quantikine Mouse IL-1ra Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse IL-1ra in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse IL-1ra and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse IL-1ra showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring mouse IL-1ra.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Mouse

Specificity

Natural and recombinant mouse IL-1ra

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

Interference observed with 1 or more available related molecules.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 26 26 27
Mean (pg/mL) 63 227 1226 66 225 1233
Standard Deviation 5.1 12.2 47.2 6.1 12.5 29.9
CV% 8.1 5.4 3.8 9.2 5.6 2.4

Recovery for Mouse IL-1ra/IL-1F3 Quantikine ELISA Kit

The recovery of mouse IL-1ra spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=5) 110 95-120
EDTA Plasma (n=4) 92 82-101
Heparin Plasma (n=4) 91 83-101
Serum (n=4) 95 88-104

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of mouse IL-1ra in each matrix were diluted with Calibrator Diluent and then assayed.

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

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

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

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

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 Quantikine ELISA Kit

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 Quantikine ELISA Kit

For research use only

Citations for Mouse IL-1ra/IL-1F3 Quantikine ELISA Kit (30)

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  • Mouse IL-1ra/IL-1F3 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: bone marrow
    Verified Customer | Posted 12/22/2021
    10 times dilution
    Mouse IL-1ra/IL-1F3 Quantikine ELISA Kit MRA00
  • Mouse IL-1ra/IL-1F3 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: EDTA Plasma and Bone marrow lavage
    Verified Customer | Posted 05/21/2021
    Mouse IL-1ra/IL-1F3 Quantikine ELISA Kit MRA00

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Protocols

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⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.