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Mouse IL-1 beta/IL-1F2 Quantikine ELISA Kit
Mouse IL-1 beta/IL-1F2 Quantikine ELISA Kit
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Mouse IL-1 beta/IL-1F2 Quantikine ELISA Kit

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Summary Sample Values Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Mouse IL-1 beta/IL-1F2 Quantikine ELISA Kit Summary

Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Tissue Lysates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
Sensitivity
4.8 pg/mL
Assay Range
12.5 - 800 pg/mL (Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma)
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.

Sample TypeMean of Detectable (pg/mL)% DetectableRange (pg/mL)
Serum (n=20)27.610ND-37.9
ND=Non-detectable

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 TypeDay 3 (pg/mL)Day 6 (pg/mL)
J774A.1 UnstimulatedNDND
J774A.1 Stimulated with LPS + GM-CSF587938
ND=Non-detectable

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)
Brain20.9
Heart28.3
Kidney111
Liver347
Lung115
Spleen987





Product Summary

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.

Precision

Intra-Assay Precision (Precision within an assay)
Inter-Assay Precision (Precision between assays)

Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma

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 8.4 6.6 5.7

Recovery

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.
Mouse IL-1 beta/IL-1F2 ELISA Linearity

Scientific Data

N/A Mouse IL-1 beta/IL-1F2 ELISA Standard Curve View Larger

Mouse IL-1 beta/IL-1F2 ELISA Standard Curve

N/A Human E‑Cadherin Antibody View Larger

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. *P< 0.05 compared with LPS alone. #P< 0.05. Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0137837), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: IL-1 beta/IL-1F2

The Interleukin 1 (IL-1) family of proteins consists of IL-1 alpha, IL-1 beta, and the IL-1 receptor antagonist (IL-1ra). IL-1 alpha and IL-1 beta bind to the same cell surface receptors and share biological functions (1). IL-1 is not produced by unstimulated cells of healthy individuals with the exception of skin keratinocytes, some epithelial cells, and certain cells of the central nervous system. However, in response to inflammatory agents, infections, or microbial endotoxins, a dramatic increase in the production of IL-1 by macrophages and various other cell types is seen. IL-1 beta plays a central role in immune and inflammatory responses, bone remodeling, fever, carbohydrate metabolism, and GH/IGF-I physiology. Inappropriate or prolonged production of IL-1 has been implicated in a variety of pathological conditions including sepsis, rheumatoid arthritis, inflammatory bowel disease, acute and chronic myelogenous leukemia, insulindependent diabetes mellitus, atherosclerosis, neuronal injury, and aging-related diseases (2-5). 

 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
Entrez Gene IDs:
3553 (Human); 16176 (Mouse); 24494 (Rat); 397122 (Porcine); 403974 (Canine); 100034237 (Equine); 100135556 (Guinea Pig)
Alternate Names:
catabolin; IL1 beta; IL-1 beta; IL-1; IL1B; IL-1b; IL1-BETA; IL-1F2; IL1F2IL-1 beta; interleukin 1, beta; interleukin-1 beta; preinterleukin 1 beta; pro-interleukin-1-beta
&#9888; 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.
背景
背景

Background: IL-1 beta/IL-1F2

The Interleukin 1 (IL-1) family of proteins consists of IL-1 alpha, IL-1 beta, and the IL-1 receptor antagonist (IL-1ra). IL-1 alpha and IL-1 beta bind to the same cell surface receptors and share biological functions (1). IL-1 is not produced by unstimulated cells of healthy individuals with the exception of skin keratinocytes, some epithelial cells, and certain cells of the central nervous system. However, in response to inflammatory agents, infections, or microbial endotoxins, a dramatic increase in the production of IL-1 by macrophages and various other cell types is seen. IL-1 beta plays a central role in immune and inflammatory responses, bone remodeling, fever, carbohydrate metabolism, and GH/IGF-I physiology. Inappropriate or prolonged production of IL-1 has been implicated in a variety of pathological conditions including sepsis, rheumatoid arthritis, inflammatory bowel disease, acute and chronic myelogenous leukemia, insulindependent diabetes mellitus, atherosclerosis, neuronal injury, and aging-related diseases (2-5). 

 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
Entrez Gene IDs:
3553 (Human); 16176 (Mouse); 24494 (Rat); 397122 (Porcine); 403974 (Canine); 100034237 (Equine); 100135556 (Guinea Pig)
Alternate Names:
catabolin; IL1 beta; IL-1 beta; IL-1; IL1B; IL-1b; IL1-BETA; IL-1F2; IL1F2IL-1 beta; interleukin 1, beta; interleukin-1 beta; preinterleukin 1 beta; pro-interleukin-1-beta
研究领域
制备和贮存
保存方式

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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货号：
MLB00C-1
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