Human IL-1ra/IL-1F3 ELISA Kit - Quantikine Best Seller
R&D Systems, part of Bio-Techne | Catalog # DRA00B
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)
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.
Cell Culture Supernates:
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.
| 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
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
Gene Symbol
IL1RN
Additional IL-1ra/IL-1F3 Products
Product Documents for Human IL-1ra/IL-1F3 Quantikine ELISA Kit
Product Specific Notices for Human IL-1ra/IL-1F3 Quantikine ELISA Kit
For research use only
⚠ 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.
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