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

R&D Systems | Catalog # DLB50

Quantikine Quickit version available - results in 90 minutes (QK201).
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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)
View all IL-1 beta/IL-1F2 ELISA Kits »

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

Species

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.

StimulantDay 1 (pg/mL)Day 3 (pg/mL)Day 5 (pg/mL)
10 μg/mL PHA218520042383
10 μg/mL PHA+10 ng/mL rhIL-2193819732839
50 ng/mL PMA176710271159
50 ng/mL LPS415821451308


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.

Human IL1B ELISA Linearity

Scientific Data Images for Human IL-1 beta/IL-1F2 Quantikine ELISA Kit

Human IL1B ELISA Cell Culture Supernate Standard Curve

Human IL1B ELISA Cell Culture Supernate Standard Curve

Human IL1B ELISA Serum/Plasma 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

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. 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, insulin dependent diabetes mellitus, atherosclerosis, neuronal injury, and aging-related diseases. 

 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

3553 (Human); 16176 (Mouse); 24494 (Rat); 397122 (Porcine); 403974 (Canine); 100034237 (Equine); 100135556 (Guinea Pig)

Gene Symbol

IL1B

Additional IL-1 beta/IL-1F2 Products

Product Documents for Human IL-1 beta/IL-1F2 Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human IL-1 beta/IL-1F2 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.

Citations for Human IL-1 beta/IL-1F2 Quantikine ELISA Kit

Customer Reviews for Human IL-1 beta/IL-1F2 Quantikine ELISA Kit (13)

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  • Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Peripheral blood mononuclear cells (PBMCs)
    Verified Customer | Posted 07/05/2024
    Human IL-1 beta/IL-1F2 Quantikine ELISA Kit DLB50
  • Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 04/24/2023
  • Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Urine
    Verified Customer | Posted 11/18/2022
    Matrix effect detected for the lowest standard in urine diluted 2-fold. Was able to detect low concentrations in urine at 2-fold dilution.
    Human IL-1 beta/IL-1F2 Quantikine ELISA Kit DLB50
  • Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 11/05/2018
  • Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 07/13/2018
  • Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell Culture Media
    Verified Customer | Posted 05/31/2018
    Human IL-1 beta/IL-1F2 Quantikine ELISA Kit DLB50
  • Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Heparin Plasma
    Verified Customer | Posted 05/08/2018
    Among best ELISAs I've worked with. Great consistency lot to lot, high sensitivity, easy to work with small amount of samples (I've diluted samples 1:100 in the past). Appropriate for concentration response curves. The only thing that can be improved is expanding the dynamic range of the assay (highest standard is at 250pg/ml and in our case we sometimes have 20,000pg/ml so need to make significant dilution to fit within the standard curve).
    Human IL-1 beta/IL-1F2 Quantikine ELISA Kit DLB50
  • Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: CHO Chinese hamster ovary cell line
    Verified Customer | Posted 04/24/2018
  • Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum and Plasma
    Verified Customer | Posted 04/04/2018
    Human IL-1 beta/IL-1F2 Quantikine ELISA Kit DLB50
  • Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: THP-1 human acute monocytic leukemia cell line
    Verified Customer | Posted 06/20/2017
    Human IL-1 beta/IL-1F2 Quantikine ELISA Kit DLB50
  • Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
    Name: Mike Ehrhardt
    Sample Tested: Serum
    Verified Customer | Posted 04/24/2017
    The Human IL-1beta Quantikine ELISA kit is used for diagnostic use.
    Human IL-1 beta/IL-1F2 Quantikine ELISA Kit DLB50
  • Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
    Name: Lamberto Torralba
    Sample Tested: Peripheral blood mononuclear cells (PBMCs)
    Verified Customer | Posted 03/15/2017
    Freshly isolated PBMCs from buffy coats stimulated with LPS (1ug/mL) for 3h together with Simvastatin. Total amount of 3 million cells/well. Volume supernatant used: 100uL. Two donors. Great production of IL1B in supernatant. Increased when statins are present as expected. Experiment used as a positive control
    Human IL-1 beta/IL-1F2 Quantikine ELISA Kit DLB50
  • Human IL-1 beta/IL-1F2 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: uman plasma
    Verified Customer | Posted 04/13/2016
    Human IL-1 beta/IL-1F2 Quantikine ELISA Kit DLB50

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Protocols

View specific protocols for Human IL-1 beta/IL-1F2 Quantikine ELISA Kit (DLB50):

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 50 µL Assay Diluent for Serum & Plasma Samples Only
  3.   For Serum & Plasma Samples: Add 50 µL of Assay Diluent to each well.
    4. 200 µL Standard, Control, or Sample
  4.   Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well.
  7.   For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
    For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  8.   Aspirate and wash 3 times.
    9. 200 µL Substrate Solution
  9.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
    10. 50 µL Stop Solution
  10. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Human IL-1 beta/IL-1F2 Quantikine ELISA Kit

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  • Q: Does the Human IL-1 beta/IL-1F2 Quantikine® ELISA kit detect pro or mature IL-1 beta?

    A: The kit has been designed to measure mature human IL-1 beta. Recombinant human Pro-IL-1 beta does not interfere with the assay but displays 6.1% cross-reactivity.

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