Human IL-1 beta/IL-1F2 QuicKit ELISA

Catalog #: QK201
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QK201

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QC266: Human IL-1 beta/IL-1F2 QuicKit Control Group 266

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

Human IL-1 beta/IL-1F2 QuicKit ELISA Summary

Assay Length

80 minutes

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)

Sensitivity

1.95 pg/mL

Assay Range

15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)

Specificity

This assay recognizes 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

No significant interference observed with available related molecules.

Sample Values

Serum/Plasma - Ten serum and plasma samples from apparently healthy volunteers were evaluated for the presence of human IL-1 beta in this assay. All samples measured less than 15.6 pg/mL. No medical histories were available for the donors used in this study.

Cell culture Supernates - Human peripheral blood mononuclear cells (PBMCs) (1 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were left unstimulated or stimulated with 10 μg/mL PHA for 24 hours. Aliquots of the culture supernates were removed, assayed for levels of human IL-1 beta, and measured at 37.2 pg/mL or 2,273 pg/mL, respectively.

PBMCs (1 x 106 cells/mL) were cultured as above. Cells were left unstimulated or stimulated with 1.0 μg/mL LPS for 24 hours. Aliquots of the cell culture supernates were removed, assayed for levels of human IL-1 beta, and were not detectable or measured 25,396 pg/mL, respectively.

Product Summary

The Quantikine^® QuicKit™ Human IL-1 beta /IL-1F2 Immunoassay is a one step, 80-minute solid phase ELISA designed to measure human 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 protein. Results obtained for naturally occurring human IL-1 beta showed linear

curves that were parallel to the standard curves obtained using the recombinant QuicKit™ standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-1 beta.

Precision

Intra-Assay Precision (Precision within an assay) Two samples of known concentration were tested twenty times on one plate to assess intra-assay precision

Inter-Assay Precision (Precision between assays) Two samples of known concentration were tested in ten separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma

	Intra-Assay Precision		Inter-Assay Precision
Sample	1	2	1	2
n	20	20	10	10
Mean (pg/mL)	102	526	103	528
Standard Deviation	1.93	7.77	4.22	16.4
CV%	1.9	1.5	4.1	3.1

Recovery

The recovery of human IL-1 beta spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.

Sample Type	Average % Recovery	Range %
Cell Culture Media (n=4)	105	99-108
EDTA Plasma (n=2)	110	106-118
Heparin Plasma (n=2)	105	99-113
Serum (n=2)	98	94-104

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human IL-1 beta in various matrices and diluted to produce samples with values within the dynamic range of the assay.

Scientific Data

N/A

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Human IL-1 beta Standard Curve

ELISA

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Human IL-1b QuicKit Spiked Recovery Competitor Comparison IL-1b is spiked at three known concentrations throughout the range of the assay and run to measure response of the spiked sample matrix. Serum recovery is 107% compared to 65% for the top competitor. EDTA plasma recovery is 111% compared to 62% for the top competitor. Heparin plasma recovery is 115% compared to 68% for the top competitor. In spike and recovery experiments, natural samples are spiked with the recombinant target analyte of interest to identify interference caused by sample matrices.

ELISA

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Human IL-1b QuicKit Linearity Competitor Comparison Samples containing and/or spiked with high concentrations of Leptin in various matrices and diluted with appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay. The linearity is between 75%-102% compared to 124%-269% for the top competitor.

Product Datasheets

Product Datasheet

COA

SDS

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 the classic members IL-1 alpha, IL-1 beta, and IL-1ra, plus IL-18, IL-33 and IL-1F5-F10. 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 observed. 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 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 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 (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

Related Research Areas

Citation for Human IL-1 beta/IL-1F2 QuicKit ELISA

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