Human IL-18 BPa 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
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: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).
Blocking Buffer: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).
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), or equivalent
Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent
Scientific Data
Product Datasheets
Preparation and Storage
Background: IL-18 BPa
Interleukin 18 binding proteins are secreted glycoproteins which antagonize IL-18 activity by binding IL-18 and blocking its biological activity. Alternative splicing of the IL-18 BP gene produces at least four isoforms that differ in affinity for IL-18. The IL-18 BP isoforms a and c each contain one immunoglobulin (Ig)-like C2-type domain while isoforms b and d lack a complete Ig-like domain.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody (to the working concentration stated in the product datasheet ) 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 each well of the microplate as recommended in the product datasheet. Incubate at room temperature for a minimum of 1 hour.
Note: The recommended Reagent Diluent typically contains 1% BSA. Some DuoSet Development Kits require alternative blocking agents, or for plates to be blocked overnight with a higher percentage of BSA, please see the product datasheet for details.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
PRECAUTION
The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.
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 (as recommended in the product datasheet), 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-18 BPa 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.
21
Citations: Showing 1 - 10
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Insights into IL-1 family cytokines in kidney allograft transplantation: IL-18BP and free IL-18 as emerging biomarkers
Authors: Cecrdlova, E;Krupickova, L;Fialova, M;Novotny, M;Tichanek, F;Svachova, V;Mezerova, K;Viklicky, O;Striz, I;
Cytokine
Species: Human
Sample Types: Serum
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Imbalanced IL-1B and IL-18 Expression in S�zary Syndrome
Authors: KCG Manfrere, MP Torrealba, FM Ferreira, ESA de Sousa, D Miyashiro, FME Teixeira, RWA Custódio, HI Nakaya, YAL Ramos, MN Sotto, A Woetmann, N Ødum, AJDS Duarte, JA Sanches, MN Sato
International Journal of Molecular Sciences, 2023-02-28;24(5):.
Species: Human
Sample Types: Serum
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Interleukin-18 signaling system links to agitation in severe mental disorders
Authors: G Hjell, A Szabo, L Mørch-John, R Holst, N Tesli, C Bell, T Fischer-Vi, MCF Werner, SH Lunding, MBEG Ormerod, IT Johansen, I Dieset, S Djurovic, I Melle, T Ueland, OA Andreassen, NE Steen, UK Haukvik
Psychoneuroendocrinology, 2022-03-12;140(0):105721.
Species: Human
Sample Types: Serum
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Interleukin-18 and Hematopoietic Recovery after Allogeneic Stem Cell Transplantation
Authors: A Radujkovic, L Kordelas, R Bogdanov, C Müller-Tid, DW Beelen, P Dreger, T Luft
Cancers (Basel), 2020-09-28;12(10):.
Species: Human
Sample Types: Serum
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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, 2019-11-14;0(0):104513.
Species: Human
Sample Types: Serum
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Early malaria infection, dysregulation of angiogenesis, metabolism and inflammation across pregnancy, and risk of preterm birth in Malawi: A cohort study
Authors: RE Elphinston, AM Weckman, CR McDonald, V Tran, K Zhong, M Madanitsa, L Kalilani-P, C Khairallah, SM Taylor, SR Meshnick, V Mwapasa, FO Ter Kuile, AL Conroy, KC Kain
PLoS Med., 2019-10-01;16(10):e1002914.
Species: Human
Sample Types: Plasma
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Adipose PTEN regulates adult adipose tissue homeostasis and redistribution via a PTEN-leptin-sympathetic loop
Authors: W Huang, NJ Queen, TB McMurphy, S Ali, L Cao
Mol Metab, 2019-09-28;30(0):48-60.
Species: Mouse
Sample Types: Serum
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Systemic inflammation is associated with malaria and preterm birth in women living with HIV on antiretrovirals and co-trimoxazole
Authors: CR McDonald, AM Weckman, AL Conroy, P Olwoch, P Natureeba, MR Kamya, DV Havlir, G Dorsey, KC Kain
Sci Rep, 2019-05-01;9(1):6758.
Species: Human
Sample Types: Plasma
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Resistance to visceral obesity is associated with increased locomotion in mice expressing an endothelial cell-specific fibroblast growth factor 1 transgene
Authors: T Keeley, A Kirov, WY Koh, V Demambro, I Bergquist, J Cotter, P Caradonna, ME Siviski, B Best, T Henderson, CJ Rosen, L Liaw, I Prudovsky, DJ Small
Physiol Rep, 2019-04-01;7(7):e14034.
Species: Human
Sample Types: Serum
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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, 2018-12-04;0(0):.
Species: Human
Sample Types: Serum
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Elevated hepatic DPP4 activity promotes insulin resistance and non-alcoholic fatty liver disease
Authors: C Baumeier, L Schlüter, S Saussentha, T Laeger, M Rödiger, SA Alaze, L Fritsche, HU Häring, N Stefan, A Fritsche, RW Schwenk, A Schürmann
Mol Metab, 2017-08-04;6(10):1254-1263.
Species: Mouse
Sample Types: Plasma
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Production and regulation of interleukin-1 family cytokines at the materno-fetal interface
Authors: LM Scott, AH Bryant, A Rees, B Down, RH Jones, CA Thornton
Cytokine, 2017-07-13;0(0):.
Species: Human
Sample Types: Tissue Culture Supernates
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Inflammatory and Angiogenic Factors at Mid-Pregnancy Are Associated with Spontaneous Preterm Birth in a Cohort of Tanzanian Women.
Authors: McDonald C, Darling A, Conroy A, Tran V, Cabrera A, Liles W, Wang M, Aboud S, Urassa W, Fawzi W, Kain K
PLoS ONE, 2015-08-06;10(8):e0134619.
Species: Human
Sample Types: Plasma
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Human papillomavirus E7 oncoprotein increases production of the anti-inflammatory interleukin-18 binding protein in keratinocytes.
Authors: Richards, Kathryn, Doble, Rosella, Wasson, Christop, Haider, Mohammed, Blair, G Eric, Wittmann, Miriam, Macdonald, Andrew
J Virol, 2014-01-29;88(8):4173-9.
Species: Human
Sample Types: Cell Culture Supernates
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Host biomarkers distinguish dengue from leptospirosis in Colombia: a case-control study.
Authors: Conroy A, Gelvez M, Hawkes M, Rajwans N, Liles W, Villar-Centeno L, Kain K
BMC Infect Dis, 2014-01-20;14(0):35.
Species: Human
Sample Types: Serum
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Levels of 4-1BB transcripts and soluble 4-1BB protein are elevated in the adipose tissue of human obese subjects and are associated with inflammatory and metabolic parameters.
Authors: Tu T, Kim C, Kang J, Nam-Goong I, Nam C, Kim E, Kim Y, Choi J, Kawada T, Goto T, Park T, Yoon Park J, Choi M, Yu R
Int J Obes (Lond), 2013-11-27;38(8):1075-82.
Species: Mouse
Sample Types: Cell Culture Supernates
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Quiescent innate response to infective filariae by human Langerhans cells suggests a strategy of immune evasion.
Authors: Boyd A, Bennuru S, Wang Y, Sanprasert V, Law M, Chaussabel D, Nutman T, Semnani R
Infect Immun, 2013-02-19;81(5):1420-9.
Species: Human
Sample Types: Cell Culture Supernates
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Impact of vitamin D supplementation on markers of inflammation in adults with cystic fibrosis hospitalized for a pulmonary exacerbation.
Eur J Clin Nutr, 2012-07-18;66(9):1072-4.
Species: Human
Sample Types: Serum
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IL-18 mediates proapoptotic signaling in renal tubular cells through a Fas ligand-dependent mechanism.
Authors: Zhang H, Hile KL, Asanuma H
Am. J. Physiol. Renal Physiol., 2011-04-20;301(1):F171-8.
Species: Human
Sample Types: Serum
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IL-8 dictates glycosaminoglycan binding and stability of IL-18 in cystic fibrosis.
Authors: Reeves EP, Williamson M, Byrne B, Bergin DA, Smith SG, Greally P, O'Kennedy R, O'Neill SJ, McElvaney NG
J. Immunol., 2009-12-21;184(3):1642-52.
Species: Human
Sample Types: Serum
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Defective phosphorylation of interleukin-18 receptor beta causes impaired natural killer cell function in systemic-onset juvenile idiopathic arthritis.
Authors: De Jager W, Vastert SJ, Beekman JM, Wulffraat NM, Kuis W, Coffer PJ, Prakken BJ
Arthritis Rheum., 2009-09-01;60(9):2782-93.
Species: Human
Sample Types: Plasma
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