Rat IL-18 DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
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
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant rat 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.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)
Product Datasheets
Preparation and Storage
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.
Assay Procedure
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.
Citations for Rat 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.
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Citations: Showing 1 - 7
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Establishment of a Rat Model of Alcoholic Liver Fibrosis with Simulated Human Drinking Patterns and Low-Dose Chemical Stimulation
Authors: Lin, CY;Omoscharka, E;Liu, Y;Cheng, K;
Biomolecules
Species: Rat
Sample Types: Tissue Homogenates
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Ameliorative effect of cheqianzi decoction on hyperuricemia and kidney injury and underlying mechanism in rats
Authors: J Meng, J Tian, Y Zhao, C Li, Y Yi, Y Zhang, J Han, L Wang, C Pan, S Liu, C Liu, F Wang, X Tang, D Wang, S Qin, A Liang
Heliyon, 2023-04-11;9(4):e15333.
Species: Rat
Sample Types: Tissue Homogenates
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Ameliorative effect of cheqianzi decoction on hyperuricemia and kidney injury and underlying mechanism in rats
Authors: J Meng, J Tian, Y Zhao, C Li, Y Yi, Y Zhang, J Han, L Wang, C Pan, S Liu, C Liu, F Wang, X Tang, D Wang, S Qin, A Liang
Heliyon, 2023;9(4):e15333.
Species: Rat
Sample Types: Tissue Homogenates
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DUSP8/TAK1 signaling mediates neuropathic pain through regulating neuroinflammation and neuron death in a spinal nerve ligation (SNL) rat model
Authors: C Liao, H Zhou, H Chen, G Cheng, S Li, F Ma, X Yang, B Xie, W Zhang
International immunopharmacology, 2022-10-21;113(0):109284.
Species: Rat
Sample Types: Cell Culture Supernates
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HMGA2 Promotes Brain Injury in Rats with Cerebral Infarction by Activating TLR4/NF-kappaB Signaling Pathway
Authors: S Huang, Z Hong, L Zhang, J Guo, Y Li, K Li
Mediators of Inflammation, 2022-08-04;2022(0):1376959.
Species: Rat
Sample Types: Tissue Homoegenates
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Endogenous tRNA-derived small RNA (tRF3-Thr-AGT) inhibits ZBP1/NLRP3 pathway-mediated cell pyroptosis to attenuate acute pancreatitis (AP)
Authors: B Sun, Z Chen, Q Chi, Y Zhang, B Gao
Journal of Cellular and Molecular Medicine, 2021-10-13;0(0):.
Species: Rat
Sample Types: Cell Culture Supernates
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MiR-199a modulates autophagy and inflammation in rats with cerebral infarction via regulating mTOR expression
Authors: J Zhou, JS Wu, Y Yan, J Li, T Ni, W Shao, JH Mei, WZ Xiong, H Wu
Eur Rev Med Pharmacol Sci, 2020-06-01;24(11):6338-6345.
Species: Rat
Sample Types: Tissue Homogenates
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LPS- stimulated rat spleen
