Human IL-1 beta/IL-1F2 ELISA Kit - Quantikine
R&D Systems, part of Bio-Techne | Catalog # DLB50
Key Product Details
Assay Length
3.5 hours or 4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (200 µL), Serum (200 µL), EDTA Plasma (200 µL), Heparin Plasma (200 µL), Citrate Plasma (200 µL)
Sensitivity
1 pg/mL
Assay Range
3.9-250 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
Product Summary for Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
The Quantikine Human IL-1 beta Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure IL-1 beta in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IL-1 beta and antibodies raised against the recombinant factor. It has been shown to quantitate recombinant human IL-1 beta accurately. Results obtained using natural IL-1 beta 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 IL-1 beta. Reports indicate that this and other ELISA kits calibrated using mature IL-1 beta as a standard will detect, but considerably underestimate, the unprocessed IL-1 beta precursor present in samples. In biological samples other than cell lysates, the precursor form of IL-1 beta is usually not the predominant form of IL-1 beta present and, additionally, is not biologically active. Therefore, results obtained using this kit should provide a useful measure of the levels of active IL-1 beta present in biological fluids.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Human
Specificity
Natural and recombinant human IL-1 beta
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 - Forty serum and plasma samples from apparently healthy volunteers were
evaluated for the presence of human IL-1 beta in this assay. No medical histories were available for
the donors used in this study. All samples measured less than the lowest IL-1 beta standard,
3.9 pg/mL.
Cell Culture Supernates - Human peripheral blood mononuclear cells (1 x 106
cells/mL) were
cultured in RPMI supplemented with 10% fetal bovine serum, 50 μM beta-mercaptoethanol,
2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were
stimulated with the agents listed in the table below. Aliquots of the cell culture supernate were
removed on days 1, 3, and 5 and assayed for levels of human IL-1 beta.
| Stimulant | Day 1 (pg/mL) | Day 3 (pg/mL) | Day 5 (pg/mL) |
| 10 μg/mL PHA | 2185 | 2004 | 2383 |
| 10 μg/mL PHA+10 ng/mL rhIL-2 | 1938 | 1973 | 2839 |
| 50 ng/mL PMA | 1767 | 1027 | 1159 |
| 50 ng/mL LPS | 4158 | 2145 | 1308 |
Precision
Intra-Assay Precision (Precision within an assay) Four 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 | 4 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 12.6 | 30.4 | 69.2 | 192 | 30.8 | 120 | 194 |
| Standard Deviation | 0.3 | 1.0 | 1.6 | 6.5 | 2.2 | 4.1 | 7.9 |
| CV% | 2.4 | 3.3 | 2.3 | 3.4 | 7.1 | 3.4 | 4.1 |
Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | ||||||
|---|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 4 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 18.9 | 30.2 | 122 | 181 | 29.8 | 118 | 193 |
| Standard Deviation | 1.6 | 1.0 | 5.4 | 5.0 | 2.5 | 5.0 | 8.0 |
| CV% | 8.5 | 3.3 | 4.4 | 2.8 | 8.4 | 4.2 | 4.1 |
Recovery for Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
The recovery of IL-1 beta spiked to different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 97 | 80-111 |
| Citrate Plasma (n=10) | 93 | 83-110 |
| EDTA Plasma (n=10) | 86 | 81-100 |
| Heparin Plasma (n=10) | 82 | 76-100 |
| Serum (n=10) | 95 | 87-110 |
Linearity
To assess linearity of the assay, the following biological samples containing, or spiked with, high concentrations of IL-1 beta were diluted with the appropriate Calibrator Diluent and then assayed.
Scientific Data Images for Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
Human IL1B ELISA Cell Culture Supernate Standard Curve
Human IL1B ELISA Serum/Plasma 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-1 beta/IL-1F2
IL-1 alpha and IL-1 beta are structurally related polypeptides that
show approximately 25% homology at the amino acid level. Both are synthesized
as 31 kDa precursors that are subsequently cleaved into mature proteins of
approximately 17.5 kDa. Cleavage of the IL-1 beta precursor by Caspase-1/ICE is a
key step in the inflammatory response. Neither IL-1 alpha nor IL-1 beta contains a
typical hydrophobic signal peptide, but evidence suggests that these factors
can be secreted by non-classical pathways. A portion of unprocessed IL-1 alpha can
be presented on the cell membrane and may retain biological activity. The
precursor form of IL-1 beta, unlike the IL-1 alpha precursor, shows little or no
biological activity in comparison to the processed form. Both unprocessed and
mature forms of IL-1 beta are exported from the cell.
IL-1 alpha and IL-1 beta exert their effects through immunoglobulin
superfamily receptors that additionally bind IL-1ra. The 80 kDa transmembrane
type I receptor (IL-1 RI) is expressed on T cells, fibroblasts, keratinocytes,
endothelial cells, synovial lining cells, chondrocytes, and hepatocytes. The 68
kDa transmembrane type II receptor (IL-1 RII) is expressed on B cells,
neutrophils, and bone marrow cells. The two IL-1 receptor types show
approximately 28% homology in their extracellular domains but differ significantly
in that the type II receptor has a cytoplasmic domain of only 29 amino acids
(aa), whereas the type I receptor has a 213 aa cytoplasmic domain. IL-1 RII
does not appear to signal in response to IL-1 and may function as a decoy
receptor that attenuates IL-1 function. The IL-1 receptor accessory protein
(IL-1 RAcP) associates with IL-1 RI and is required for IL-1 RI signal
transduction. IL-1ra is a secreted molecule that functions as a competitive
inhibitor of IL-1. Soluble forms of both IL-1 RI and IL-1 RII have been
detected in human plasma, synovial fluids, and the conditioned media of several
human cell lines. In addition, IL-1 binding proteins that resemble soluble IL-1
RII are encoded by vaccinia and cowpox viruses.
Long Name
Interleukin 1 beta
Alternate Names
IL-1b, IL-1F2, IL1 beta, IL1B
Entrez Gene IDs
Gene Symbol
IL1B
Additional IL-1 beta/IL-1F2 Products
Product Documents for Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
Product Specific Notices for Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
For research use only
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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.