Human IL-1ra/IL-1F3 ELISA Kit - Quantikine Best Seller

Name: Human IL-1ra/IL-1F3 Quantikine ELISA Kit
Brand: R&D Systems
SKU: DRA00B
Rating: 5 (10 reviews)

R&D Systems, part of Bio-Techne | Catalog # DRA00B

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Product Summary
Product Specifications
Sample Values
Precision
Recovery
Linearity
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (100 µL), Serum (100 µL), EDTA Plasma (100 µL), Heparin Plasma (100 µL)

Sensitivity

18.3 pg/mL

Assay Range

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

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

The Quantikine Human IL-1ra Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-1ra in cell culture supernates, serum, and plasma. It contains E. coli-derived recombinant human IL-1ra as well as antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human IL-1ra. Results obtained using natural human 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 natural human IL-1ra.

Product Specifications

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Human

Specificity

Natural and recombinant human 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.

Sample Values

Serum/Plasma - Samples from apparently healthy volunteers were evaluated for the presence of human IL-1ra in this assay. No medical histories were available for the donors used in this study.

Sample Type	Mean (pg/mL)	Range (pg/mL	Standard Deviation (pg/mL)
Serum (n=35)	360	181-1327	214
EDTA plasma (n=35)	230	105-1062	177
Heparin plasma (n=35)	309	160-1193	194

Cell Culture Supernates:

Human peripheral blood cells (1 x 106 cells/mL) were cultured in DMEM supplemented with 5% fetal bovine serum, 50 μM beta-mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 10 μg/mL PHA. Aliquots of the cell culture supernates were removed and assayed for levels of human IL-1ra.

Condition	Day 1 (pg/mL)	Day 6 (pg/mL)
Unstimulated	1286	49,621
Stimulated	9998	18,803

Jurkat human acute T cell leukemia cells (1 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum. An aliquot of the cell culture supernate was removed and assayed for human IL-1ra. No detectable levels were observed.

IMR-90 human lung fibroblast cells (1 x 106 cells/mL) were cultured in DMEM supplemented with 10% fetal bovine serum. An aliquot of the cell culture supernate was removed and assayed for human IL-1ra. No detectable levels were observed.

THP-1 human acute monocytic leukemia cells (1 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum and 50 μM beta-mercaptoethanol. An aliquot of the cell culture supernate was removed, assayed for human IL-1ra, and measured 1464 pg/mL.

U937 human histiocytic lymphoma cells (1 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum and 2 mM L-glutamine. An aliquot of the cell culture supernate was removed, assayed for human IL-1ra, and measured 134 pg/mL.

HEK293 human embryonic kidney cells (1 x 106 cells/mL) were cultured in DMEM supplemented with 10% fetal bovine serum. An aliquot of the cell culture supernate was removed after 5 days and assayed for human IL-1ra. No detectable levels were observed.

HepG2 human hepatocellular carcinoma cells (1 x 106 cells/mL) were cultured in DMEM supplemented with 10% fetal bovine serum. An aliquot of the cell culture supernate was removed, assayed for human IL-1ra, and measured 597 pg/mL.

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

	Intra-Assay Precision			Inter-Assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	40	40	40
Mean (pg/mL)	64.9	538	1343	56.2	469	1243
Standard Deviation	3.67	22.1	55.7	5.79	31.9	70.4
CV%	5.6	4.1	4.1	10.3	6.8	5.7

EDTA Plasma, Heparin Plasma, Serum

	Intra-Assay Precision			Inter-Assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	40	40	40
Mean (pg/mL)	66.9	607	1627	82.5	646	1663
Standard Deviation	4.88	30.2	60.0	9.04	51.4	111
CV%	7.3	5.0	3.7	11.0	8.0	6.7

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

The recovery of 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=4)	106	96-115
EDTA Plasma (n=4)	86	66-102
Heparin Plasma (n=4)	93	82-106
Serum (n=4)	90	81-99

Linearity

To assess the linearity of the assay, samples containing high concentrations of IL-1ra were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.

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

Human IL-1ra ELISA/IL-1F3 ELISA Calibrator Diluent RD5-33 (1X) Standard Curve

Human IL-1ra ELISA/IL-1F3 ELISA Calibrator Diluent RD6-3 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

Interleukin-1 receptor antagonist (IL-1ra; also known as IL-1F3) is a 22-25 kDa member of the IL-1 family of cytokines. Currently, there are 11 family members (IL-1F1-F11), nine of which form an IL-1 gene cluster on human Ch2 (1-3). Each IL-1 family member contains an IL-1 fold. This fold is generated by 12 packed beta-sheets that interact to form a beta-trefoil structure. Little amino acid (aa) homology is required to achieve this structure, and this explains the low aa identity among IL-1 family members. IL-1ra is a pure cytokine receptor antagonist that has no signal transduction-initiating activity (4). It is an acute phase protein that exists to dampen inflammation. IL-1( beta) is initially produced by monocytes in response to a variety of stimuli. Circulating IL-1 then binds to widely expressed IL-1 type I receptors (IL-1 RI) and initiates a number of pro-inflammatory events. On endothelial cells (EC), IL-1 induces PGE2 and IL-6 release, generating fever, thrombocytosis, and hepatic acute phase protein production. In synovial joints, IL-1 induces chondrocyte NO production, an event that leads to reduced collagen synthesis and chondrocyte apoptosis. Finally, IL-1 increases neutrophil counts, both in blood and tissue, and thus is able to promote a pro-inflammatory environment in multiple locations (5-8). IL-1ra blocks IL-1 action through competitive inhibition. More correctly, although IL-1ra fills the IL-1 binding site in IL-1 RI, it is also unable to orchestrate the creation of a signal-transducing IL-1 RI:IL-1 R Accessory protein (IL-1 R AcP) heterodimer complex. Effective IL-1ra concentrations are generally 100-fold greater than local IL-1 concentrations. This is because the IL-1ra half-life is but 6 minutes, and very few IL-1 type I receptors need to be engaged by IL-1 to elicit a cellular response (5, 7, 9).

Human IL-1ra is synthesized as a 177 aa precursor that contains a 25 aa signal sequence and a 152 aa mature region (10, 11). Although it contains an IL-1 cytokine fold, it apparently lacks two structural motifs that allow for activation of the IL-1 receptor heterodimer. First, and following binding to IL-1 RI, the presence of Ile 51-His 54 and Lys 145 of the mature molecule preclude recruitment of IL-1 R AcP. Second, there is no identifiable C-terminal lectin segment that is hypothesized to help recruit an accessory signaling component (1, 12, 13). Mature human IL-1ra is 77% and 82% aa identical to mouse and canine IL-1ra, respectively, and human IL-1ra inhibits IL-1 activity on mouse cells (10). A number of cell types express IL-1ra, including monocytes (11), Sertoli cells (14), hepatocytes (15), adipocytes (16), synovial fibroblasts (17), mast cells (18), pancreatic beta-cells (19), and intestinal epithelial cells (20). There are at least three intracellular IL-1ra isoforms (icIL-1ra1, 2, and 3). All show N-terminal variation, and all contain amino acids 35-177 of the secreted precursor (21-23). Intracellular IL-1ra1 is of particular interest, because it is reported to be "secreted" by endothelial cells and binds to the IL-1 RI in an antagonist fashion (23-25). Intracellular IL-1ra1 is 159 aa in length and shows a 3 aa substitution for the first 21 aa's of the signal sequence of IL-1ra (21).

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

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Product Specific Notices for Human IL-1ra/IL-1F3 Quantikine ELISA Kit

For research use only

Citations
Reviews

⚠ 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.