Human IL-38/IL-1F10 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
- 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)
Scientific Data
Product Datasheets
Preparation and Storage
Background: IL-38/IL-1F10
Human IL-38/IL-1F10 is a secreted cytokine belonging to the IL-1 family. It is synthesized as a 152 amino acid mature protein that lacks an identifiable signal sequence. It binds to soluble IL-1 R1. Human IL-38 shares 82% aa sequence identity with mouse IL-1F10. Human IL-38 also shares 39% and 27% aa sequence identity with IL-1ra and IL-1 beta, respectively.
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 of Block Buffer 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 Human IL-38/IL-1F10 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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Effect of exogenous IL-37 on immune cells from a patient carrying a potential IL37 loss-of-function variant: A case study
Authors: LU Teufel, CI van der Ma, V Klück, A Simons, A Hoischen, V Vernimmen, LAB Joosten, RJW Arts
Cytokine, 2022-12-05;162(0):156102.
Species: Human
Sample Types: Plasma
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Plant-produced recombinant cytokines IL-37b and IL-38 modulate inflammatory response from stimulated human PBMCs
Authors: I Kolotilin
Scientific Reports, 2022-11-14;12(1):19450.
Species: Human
Sample Types: Protein
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Association between IL-38 and inflammatory indicators in patients with bacterial pneumonia
Authors: X Sun, J Zhou, W Huang, B Wang, Y Zhang, L Duan, W Chen
Cytokine, 2022-11-12;161(0):156052.
Species: Human
Sample Types: Serum
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Interleukin 38 alleviates aortic valve calcification by inhibition of NLRP3
Authors: E The, DM de Graaf, Y Zhai, Q Yao, L Ao, DA Fullerton, CA Dinarello, X Meng
Proceedings of the National Academy of Sciences of the United States of America, 2022-08-29;119(36):e2202577119.
Species: Human
Sample Types: Cell Culture Supernates
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Serum interleukin-38 levels correlated with insulin resistance, liver injury and lipids in non-alcoholic fatty liver disease
Authors: J Cao, L Hua, S Zhang, J Tang, F Ke, Z Wu, G Xue
Lipids in Health and Disease, 2022-08-10;21(1):70.
Species: Human
Sample Types: Serum
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Reduced concentrations of the B cell cytokine Interleukin 38 are associated with cardiovascular disease risk in overweight subjects
Authors: DM de Graaf, M Jaeger, ICL van den Mu, RT Horst, K Schraa, J Zwaag, M Kox, M Fujita, T Yamauchi, L Mercurio, S Madonna, JHW Rutten, J de Graaf, NP Riksen, FL van de Vee, MG Netea, LAB Joosten, CA Dinarello
Eur J Immunol, 2020-11-19;0(0):.
Species: Human
Sample Types: Plasma
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IL-38 inhibits microglial inflammatory mediators and is decreased in amygdala of children with autism spectrum disorder
Authors: I Tsilioni, H Pantazopou, P Conti, SE Leeman, TC Theoharide
Proc. Natl. Acad. Sci. U.S.A., 2020-06-29;0(0):.
Species: Human
Sample Types: Serum
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Interleukin-38 interacts with destrin/actin-depolymerizing factor in human keratinocytes
Authors: D Talabot-Ay, L Mermoud, J Borowczyk, J Drukala, M Wolnicki, A Modarressi, WH Boehncke, N Brembilla, G Palmer
PLoS ONE, 2019-11-26;14(11):e0225782.
Species: Human
Sample Types: Cell Culture Supernates
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The role of IL-36? and its regulation in eosinophilic inflammation in allergic rhinitis
Authors: X Qin, M Liu, S Zhang, C Wang, T Zhang
Cytokine, 2019-03-01;117(0):84-90.
Species: Human
Sample Types: Serum
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