Mouse IL-1 beta/IL-1F2 ELISA Kit - Quantikine Best Seller
R&D Systems, part of Bio-Techne | Catalog # MLB00C
Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Tissue Lysates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)
Sensitivity
4.8 pg/mL
Assay Range
12.5-800 pg/mL (Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma)
Product Summary for Mouse IL-1 beta/IL-1F2 Quantikine ELISA Kit
The Quantikine® Mouse IL-1 beta Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse IL-1 beta in cell culture supernates, tissue lysates, serum, and plasma. It contains E. coli-expressed recombinant mouse IL-1 beta and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural mouse IL-1 beta showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse IL-1 beta.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Mouse
Specificity
Natural and recombinant mouse IL-1 beta
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Serum/Plasma - Twenty samples were evaluated for the presence of mouse IL-1 beta in this assay.
Serum samples measured as shown in the table below. All twenty plasma samples measured
less than the lowest Mouse IL-1 beta Standard, 12.5 pg/mL.
ND=Non-detectable
ND=Non-detectable
| Sample Type | Mean of Detectable (pg/mL) | % Detectable | Range (pg/mL) |
| Serum (n=20) | 27.6 | 10 | ND-37.9 |
Cell Culture Supernates - J774A.1 mouse reticulum cell sarcoma macrophage cells
(3.5 x 106
cells/mL) were cultured in DMEM supplemented with 10% fetal bovine serum and
stimulated with 5 μM beta-mercapthoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and
100 μg/mL streptomycin sulfate. The cell culture supernate was unstimulated or stimulated
with 2.5 μg/mL of lipopolysacharride (LPS) and 100 ng/mL of recombinant mouse GM-CSF
(R&D Systems, Catalog # 415-ML) for 3 and 6 days. Aliquots of the cell culture supernates were
removed and assayed for levels of mouse IL-1 beta.
| Sample Type | Day 3 (pg/mL) | Day 6 (pg/mL) |
| J774A.1 Unstimulated | ND | ND |
| J774A.1 Stimulated with LPS + GM-CSF | 587 | 938 |
Tissue Lysates - Organs from 2-3 mice were rinsed with PBS to remove excess blood, chopped
into 1-2 mm pieces, homogenized with a tissue homogenizer, and 1 mL of Lysis Buffer 2 was
added (2 mL of Lysis Buffer 2 was added to liver tissue). Organs were lysed at room temperature
for 30 minutes with gentle agitation and centrifuged to remove debris. An aliquot of each
tissue lysate was removed and assayed for levels of mouse IL-1 beta.
| Tissue Lysate | (pg/mL) |
| Brain | 20.9 |
| Heart | 28.3 |
| Kidney | 111 |
| Liver | 347 |
| Lung | 115 |
| Spleen | 987 |
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, Tissue Lysates
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 33.5 | 92.9 | 386 | 34.6 | 93.5 | 423 |
| Standard Deviation | 2.5 | 4.3 | 11.7 | 2.9 | 6.2 | 23.9 |
| CV% | 7.5 | 4.6 | 3.0 | 8.4 | 6.6 | 5.7 |
Recovery for Mouse IL-1 beta/IL-1F2 Quantikine ELISA Kit
The recovery of mouse IL-1 beta 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 | 95-119 |
| EDTA Plasma (n=4) | 108 | 101-117 |
| Heparin Plasma (n=4) | 97 | 87-114 |
| Serum (n=4) | 105 | 98-113 |
| Tissue Lysates (n=4) | 115 | 110-120 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse IL-1 beta were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Mouse IL-1 beta/IL-1F2 Quantikine ELISA Kit
Mouse IL-1 beta/IL-1F2 ELISA Standard Curve
Human E‑Cadherin Antibody
PYC suppressed LPS-induced proinflammatory cytokine production in BV2 microglia.Cells were incubated with the indicated concentrations of PYC or vehicle for 1h before LPS treatment (500ng/mL). After 24 h incubation, the culture supernatants were collected, and the amount of TNF-alpha, IL-6, IL-1 beta and IL-10 were measured by ELISA (A-D). Data are represented as mean ± SD from at least 3 independent experiments. *PPreparation 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 (aa) level. Both are
synthesized as 31 kDa precursors that are subsequently cleaved into mature
proteins of approximately 17.5 kDa (6, 7). Cleavage of the IL-1 beta precursor by
Caspase-1/ICE is a key step in the inflammatory response (2, 8). Neither IL-1 alpha
nor IL-1 beta contains a typical hydrophobic signal peptide (9-11), but evidence
suggests that these factors can be secreted by non-classical pathways (12, 13).
A portion of unprocessed IL-1 alpha can be presented on the cell membrane and may
retain biological activity (14). 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 (13, 15). 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 (16,
17). The 68 kDa transmembrane type II receptor (IL-1 RII) is expressed on B cells,
neutrophils, and bone marrow cells (18). 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
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 (19). The IL-1 receptor accessory protein (IL-1
RAcP) associates with IL-1 RI and is required for IL-1 RI signal transduction
(20). IL-1ra is a secreted molecule that functions as a competitive inhibitor
of IL-1 (21, 22). 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 (23, 24). In addition, IL-1 binding proteins that resemble soluble
IL-1 RII are encoded by vaccinia and cowpox viruses (25).
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 Mouse IL-1 beta/IL-1F2 Quantikine ELISA Kit
Product Specific Notices for Mouse 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.