Human Total IL-18 DuoSet ELISA

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Ancillary Products Available
Human Total IL-18 / IL-1F4 ELISA Standard Curve
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Product Details
Citations (9)
Supplemental Products
Reviews (6)

Human Total IL-18 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sufficient Materials
For five 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Total Interleukin 18 (Total IL-18). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Data Example

Human Total IL-18 / IL-1F4 ELISA Standard Curve

Product Datasheets

Preparation and Storage

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-18/IL-1F4

Interleukin 18 (IL-18), also known as interferon-gamma-inducing factor (IGIF) and IL-1γ, is a cytokine which shares biologic activities with IL-12 and structural similarities with the IL-1 family of proteins. IL-18 was originally cloned from liver cells and has since been shown to be expressed by monocyte/macrophages, osteoblasts and keratinocytes. Caspase-1 (IL-1 beta-converting enzyme) has been implicated in the physiological processing of pro-IL-18 to IL-18. Similarly to IL-12, human IL-18 has been shown to enhance NK cell activity in PBMC cultures. Human IL-18 has also been found to induce the production IFN-γ and GM-CSF while inhibiting the production of IL-10 by PBMC. On enriched human T cells, human IL-18 can enhance Th1 cytokine production and stimulate cell proliferation via an IL-2-dependent pathway. In the mouse system, IL-18 has been shown to be a costimulatory factor for the activation of Th1, but not Th2, cells. IL-18 was found to selectively enhance the FasL-mediated cytotoxicity of Th1, but not Th0 or Th2, cells. IL-18 has also been shown to induce activated B cells to produce IFN-γ that inhibits IgE production.

Long Name:
Interleukin 18
Entrez Gene IDs:
3606 (Human); 16173 (Mouse); 29197 (Rat); 397057 (Porcine); 574151 (Primate)
Alternate Names:
Iboctadekin; IFN-gamma-inducing factor; IGIF; IGIFIL-1 gamma; IL18; IL-18; IL-18MGC12320; IL-1F4; IL1F4iboctadekin; IL-1g; Interferon gamma-inducing factor; interleukin 18 (interferon-gamma-inducing factor); Interleukin-1 gamma; interleukin-18

Assay Procedure


Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human Total IL-18 DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

9 Citations: Showing 1 - 9
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  1. Impact of constitutional TET2 haploinsufficiency on molecular and clinical phenotype in humans
    Authors: E Kaasinen, O Kuismin, K Rajamäki, H Ristolaine, M Aavikko, J Kondelin, S Saarinen, DG Berta, R Katainen, EAM Hirvonen, A Karhu, A Taira, T Tanskanen, A Alkodsi, M Taipale, E Morgunova, K Franssila, R Lehtonen, M Mäkinen, K Aittomäki, A Palotie, MI Kurki, O Pietiläine, M Hilpert, E Saarentaus, J Niinimäki, J Junttila, K Kaikkonen, P Vahteristo, RC Skoda, MRJ Seppänen, KK Eklund, J Taipale, O Kilpivaara, LA Aaltonen
    Nat Commun, 2019;10(1):1252.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Human metapneumovirus activates NOD-like receptor protein 3 inflammasome via its small hydrophobic protein which plays a detrimental role during infection in mice
    Authors: VB Lê, J Dubois, C Couture, MH Cavanagh, O Uyar, A Pizzorno, M Rosa-Calat, MÈ Hamelin, G Boivin
    PLoS Pathog., 2019;15(4):e1007689.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Increased interleukin 18 activity in adolescents with early-onset psychosis is associated with cortisol and depressive symptoms
    Authors: K Wedervang-, S Friis, V Lonning, RE Smelror, C Johannesse, EJ Reponen, SH Lyngstad, T Lekva, P Aukrust, T Ueland, OA Andreassen, I Agartz, AM Myhre
    Psychoneuroendocrinology, 2019;0(0):104513.
    Species: Human
    Sample Types: Serum
  4. EASIX for prediction of survival in lower-risk myelodysplastic syndromes
    Authors: A Merz, U Germing, G Kobbe, J Kaivers, A Jauch, A Radujkovic, M Hummel, A Benner, M Merz, P Dreger, T Luft
    Blood Cancer J, 2019;9(11):85.
    Species: Human
    Sample Types: Plasma
  5. Lysosomotropic beta blockers induce oxidative stress and IL23A production in Langerhans cells
    Authors: G Müller, C Lübow, G Weindl
    Autophagy, 2019;0(0):1-16.
    Species: Human
    Sample Types: Cell Culture Supernatants
  6. TET-2 up-regulation is associated with the anti-inflammatory action of Vicenin-2
    Authors: N Hassan, A Ali, C Withycombe, M Ahluwalia, RH Al-Nasseri, A Tonks, K Morris
    Cytokine, 2018;108(0):37-42.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Asymmetric dimethylarginine serum levels are associated with early mortality after allogeneic stem cell transplantation
    Authors: A Radujkovic, H Dai, L Kordelas, D Beelen, SP Rachakonda, C Müller-Tid, R Kumar, P Dreger, T Luft
    Haematologica, 2018;0(0):.
    Species: Human
    Sample Types: Serum
  8. Altered distribution of peripheral blood dendritic cell subsets in patients with pulmonary paracoccidioidomycosis
    Authors: J Venturini, RS Cavalcante, DV Moris, M de Assis G, AD Levorato, KH Dos Reis, MSP de Arruda, RP Mendes
    Acta Trop., 2017;0(0):.
    Species: Human
    Sample Types: Serum
  9. TB lymphadenitis is associated with enhanced baseline and antigen - specific induction of Type 1 and Type 17 cytokines and reduced IL-1? and IL-18 at the site of infection
    Authors: GR Kathamuthu, K Moideen, D Baskaran, VV Banurekha, D Nair, G Sekar, R Sridhar, B Vidyajayan, G Gajendrara, DK Parandhama, A Srinivasan, S Babu
    Clin. Vaccine Immunol, 2017;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates


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Reviews for Human Total IL-18 DuoSet ELISA

Average Rating: 5 (Based on 6 Reviews)

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Human Total IL-18 DuoSet ELISA
By Anonymous on 06/11/2020
Application: Sample Tested: Cell culture supernatant,THP-1 cells

The PMA-differentiated THP-1 cells were treated with or without LPS/ATP for 4 h. The production of IL-18 was tested from culture supernatants of treated THP-1 cells.

Human Total IL-18 DuoSet ELISA
By Anonymous on 11/13/2019
Application: Sample Tested: THP-1 human acute monocytic leukemia cell line,Cell culture supernatant

Human Total IL-18 DuoSet ELISA
By Anonymous on 10/02/2018
Application: Sample Tested: Serum and Plasma

Human Total IL-18 DuoSet ELISA
By Anonymous on 02/06/2018
Application: Sample Tested: Bone marrow-derived dendritic cells

Human Total IL-18 DuoSet ELISA
By Anonymous on 11/21/2017
Application: Sample Tested: Serum

We used human serum at a dilution of 1:10. The result was very good.

Human Total IL-18 DuoSet ELISA
By Anonymous on 05/17/2017
Application: Sample Tested: human serum