IL-18 (Interleukin-18) is secreted by macrophages, dendritic cells, and epithelial cells. Circulating mature IL-18 is sequestered by soluble IL-18 binding proteins (IL-18 BP) that inhibit IL-18 bioactivity. IL-18 signals through a receptor complex that contains IL-18 R alpha IL-18 R beta. It synergizes with other cytokines to activate NK, Th1, and Th17 cells and to increase the production of IFN-gamma. It can also promote Th2 cytokine release which reduces the effectiveness of antiviral responses. Increased levels of active IL-18 contribute to the severity of autoimmunity and hypertension, while deficiency of IL-18 results in symptoms of metabolic syndrome. In cancer, IL-18 stimulates Th1 and NK cells to target tumor cells, but it can also promote angiogenesis, metastasis, and tumor cell immune evasion.
Human Total IL-18 DuoSet ELISA
R&D Systems | Catalog # DY318-05
Key Product Details
Assay Type
Assay Range
Sample Type
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Reactivity
Human Total IL-18 DuoSet ELISA 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
Product Summary for Human Total IL-18 DuoSet ELISA
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 Specifications
Assay Format
Detection Method
Conjugate
Specificity
Label
Scientific Data Images for Human Total IL-18 DuoSet ELISA
Human Total IL-18 / IL-1F4 ELISA Standard Curve
Kit Contents for Human Total IL-18 DuoSet ELISA
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008C) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
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 equivalent
Reagent Diluent*
Blocking Buffer*
Substrate Solution: ELISA TMB Substrate (Catalog # DY999B or DY999B-250)
Stop Solution: Methanesulfonic acid (Catalog # DY994B or DY994B-250)
Microplates: (Catalog # DY990), or equivalent
Plate Sealers: (Catalog # DY992), or equivalent
*For the recommended Reagent Diluent and Blocking Buffer for a specific DuoSet ELISA Development Kit, refer to the product datasheet.
Preparation and Storage
Shipping
Stability & Storage
Background: IL-18/IL-1F4
Long Name
Alternate Names
Gene Symbol
Additional IL-18/IL-1F4 Products
Product Documents for Human Total IL-18 DuoSet ELISA
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 Total IL-18 DuoSet ELISA
For research use only
Citations for Human Total IL-18 DuoSet ELISA
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Customer Reviews for Human Total IL-18 DuoSet ELISA (10)
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Customer Images
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Sample Tested: Cancer cell lysatesVerified Customer | Posted 02/24/2023The kit was essential to quantify this cytokine in Caco-2 cells, previously grown for 21 days and then treated with pro-inflammatory factors (LPS and IFN-gamma). Results obtained agreed with other reports in the literature and were highly replicable.
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Sample Tested: Serum-free Cell Culture SupernatesVerified Customer | Posted 02/14/2023HLF cells treated with LPS+ATP
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Sample Tested: SerumVerified Customer | Posted 10/05/2020
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Sample Tested: Tissue Culture MediaVerified Customer | Posted 09/29/2020Detection of IL-18 produced by human CAR-T cells engineered to express IL-18 (PMC659), compared to normal CAR-T cells (PMC650).
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Sample Tested: Cell culture supernatant and THP-1 cellsVerified Customer | Posted 06/11/2020The 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.
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Sample Tested: THP-1 human acute monocytic leukemia cell line and Cell culture supernatantVerified Customer | Posted 11/13/2019
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Sample Tested: Serum and PlasmaVerified Customer | Posted 10/02/2018
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Sample Tested: Bone marrow-derived dendritic cellsVerified Customer | Posted 02/06/2018
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Sample Tested: SerumVerified Customer | Posted 11/21/2017We used human serum at a dilution of 1:10. The result was very good.
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Sample Tested: Human serumVerified Customer | Posted 05/17/2017
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Protocols
View specific protocols for Human Total IL-18 DuoSet ELISA (DY318-05):
GENERAL ELISA PROTOCOL
Plate Preparation
- 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.
- 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.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- 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.
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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