Human IL-13 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.
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human IL-13. The suggested diluent is suitable for the analysis of most cell culture supernate 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)
Scientific Data
Product Datasheets
Preparation and Storage
Background: IL-13
IL-13 (Interleukin-13) plays a key role in the pathogenesis of allergy, cancer, and tissue fibrosis. It is secreted by several helper T cell subsets, NK cells, mast cells, eosinophils, basophils, and visceral smooth muscle cells. IL-13 suppresses the production of proinflammatory cytokines and other cytotoxic substances by macrophages, fibroblasts, and endothelial cells. It promotes B cell activation, immunoglobulin class switching to IgE, and the upregulation of CD23/Fc epsilon RII. IL-13 signals through a receptor complex containing IL-13 R alpha 1 and IL-4 R alpha. This complex also functions as the type 2 IL-4 receptor. IL-13 R alpha 2 binds IL-13 and prevents IL-13 from signaling through the IL-13 R alpha 1/IL-4 R alpha complex. IL-13-bound IL-13 R alpha 2 can directly promote tumor cell invasiveness and the development of tissue fibrosis.
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 Human IL-13 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 - 10
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Suppression of NK Cell Activation by JAK3 Inhibition: Implication in the Treatment of Autoimmune Diseases
Authors: Wu, WC;Shiu, C;Tong, TK;Leung, SO;Hui, CW;
Journal of immunology research
Species: Human
Sample Types: Cell Culture Supernates
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The Functional DNA Methylation Signatures Relevant to Altered Immune Response of Neonatal T Cells with l-Arginine Supplementation
Authors: HR Yu, TY Hsu, CC Tsai, HC Huang, HH Cheng, YJ Lai, YJ Lin, CC Chen, SC Li, K Yang
Nutrients, 2021-08-13;13(8):.
Species: Human
Sample Types: Cell Culture Supernates
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Bacterial genotoxins induce T�cell senescence
Authors: SL Mathiasen, L Gall-Mas, IS Pateras, SDP Theodorou, MRJ Namini, MB Hansen, OCB Martin, CK Vadivel, K Ntostoglou, D Butter, M Givskov, C Geisler, AN Akbar, VG Gorgoulis, T Frisan, N Ødum, T Krejsgaard
Cell Reports, 2021-06-08;35(10):109220.
Species: Human
Sample Types: Cell Culture Supernates
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Monocytes differentiated into macrophages and dendritic cells in the presence of human IFN-&lambda3 or IFN-&lambda4 show distinct phenotypes
Authors: M De, A Bhushan, S Chinnaswam
J Leukoc Biol, 2020-11-17;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Gaps in Study Design for Immune Parameter Research for Latent Tuberculosis Infection: A Systematic Review
Authors: M Herrera, C Vera, Y Keynan, ZV Rueda
J Immunol Res, 2020-04-21;2020(0):8074183.
Species: Human
Sample Types: Cell Culture Supernates
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Identification of a novel role of IL-13R?2 in human Glioblastoma multiforme: interleukin-13 mediates signal transduction through AP-1 pathway
Authors: R Bhardwaj, A Suzuki, P Leland, BH Joshi, RK Puri
J Transl Med, 2018-12-20;16(1):369.
Species: Human
Sample Types: Whole Cells
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Phosphatase of regenerating liver-3 (PRL-3) is overexpressed in classical Hodgkin lymphoma and promotes survival and migration
Authors: MA Hjort, H Hov, P Abdollahi, EN Vandsemb, UM Fagerli, B Lund, TS Slørdahl, M Børset, TB Rø
Exp Hematol Oncol, 2018-04-10;7(0):8.
Species: Human
Sample Types: Cell Culture Supernates
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A Proinflammatory Role of Type 2 Innate Lymphoid Cells in Murine Immune-Mediated Hepatitis
Authors: Gisa Tiegs
J. Immunol., 2016-11-21;0(0):.
Species: Human
Sample Types: Plasma
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Characterization of host and microbial determinants in individuals with latent tuberculosis infection using a human granuloma model.
Authors: Guirado E, Mbawuike U, Keiser T, Arcos J, Azad A, Wang S, Schlesinger L
MBio, 2015-02-17;6(1):e02537-14.
Species: Human
Sample Types: Cell Culture Supernates
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Immunoepidemiological profiling of onchocerciasis patients reveals associations with microfilaria loads and ivermectin intake on both individual and community levels.
Authors: Arndts, Kathrin, Specht, Sabine, Debrah, Alexande, Tamarozzi, Francesc, Klarmann Schulz, Ute, Mand, Sabine, Batsa, Linda, Kwarteng, Alexande, Taylor, Mark, Adjei, Ohene, Martin, Coralie, Layland, Laura E, Hoerauf, Achim
PLoS Negl Trop Dis, 2014-02-20;8(2):e2679.
Species: Human
Sample Types: Cell Culture Supernates
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The aryl hydrocarbon receptor is functionally upregulated early in the course ofvhuman T-cell activation.
Authors: Prigent L, Robineau M, Jouneau S, Morzadec C, Louarn L, Vernhet L, Fardel O, Sparfel L
Eur J Immunol, 2014-02-18;44(5):1330-40.
Species: Human
Sample Types: Cell Culture Supernates
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CD137 ligand signalling induces differentiation of primary acute myeloid leukaemia cells.
Authors: Cheng K, Wong S, Linn Y, Ho L, Chng W, Schwarz H
Br J Haematol, 2014-01-15;165(1):134-44.
Species: Human
Sample Types: Cell Culture Supernates
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CD137L-stimulated dendritic cells are more potent than conventional dendritic cells at eliciting cytotoxic T-cell responses.
Authors: Harfuddin Z, Kwajah S, Chong Nyi Sim A, Macary P, Schwarz H
Oncoimmunology, 2013-11-11;2(11):e26859.
Species: Human
Sample Types: Cell Culture Supernates
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Interleukin-33-dependent innate lymphoid cells mediate hepatic fibrosis.
Authors: McHedlidze T, Waldner M, Zopf S, Walker J, Rankin A, Schuchmann M, Voehringer D, McKenzie A, Neurath M, Pflanz S, Wirtz S
Immunity, 2013-08-15;39(2):357-71.
Species: Human
Sample Types: Serum
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Elevated adaptive immune responses are associated with latent infections of Wuchereria bancrofti.
Authors: Arndts K, Deininger S, Specht S, Klarmann U, Mand S, Adjobimey T, Debrah AY, Batsa L, Kwarteng A, Epp C, Taylor M, Adjei O, Layland LE, Hoerauf A
PLoS Negl Trop Dis, 2012-04-03;6(4):e1611.
Species: Human
Sample Types: Cell Culture Supernates
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Interferon-beta treatment in multiple sclerosis attenuates inflammatory gene expression through inducible activity of the phosphatase SHP-1.
Authors: Christophi GP, Panos M, Hudson CA, Tsikkou C, Mihai C, Mejico LJ, Jubelt B, Massa PT
Clin. Immunol., 2009-06-25;133(1):27-44.
Species: Human
Sample Types: Plasma
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Fungal proteases induce Th2 polarization through limited dendritic cell maturation and reduced production of IL-12.
Authors: Lamhamedi-Cherradi SE, Martin RE, Ito T, Kheradmand F, Corry DB, Liu YJ, Moyle M
J. Immunol., 2008-05-01;180(9):6000-9.
Species: Human
Sample Types: Cell Culture Supernates
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Diminished frequency and function of CD4+CD25high regulatory T cells associated with active uveitis in Vogt-Koyanagi-Harada syndrome.
Authors: Chen L, Yang P, Zhou H, He H, Ren X, Chi W, Wang L, Kijlstra A
Invest. Ophthalmol. Vis. Sci., 2008-04-17;49(8):3475-82.
Species: Human
Sample Types: Cell Culture Supernates
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Tumor-associated leukemia inhibitory factor and IL-6 skew monocyte differentiation into tumor-associated macrophage-like cells.
Authors: Duluc D, Delneste Y, Tan F, Moles MP, Grimaud L, Lenoir J, Preisser L, Anegon I, Catala L, Ifrah N, Descamps P, Gamelin E, Gascan H, Hebbar M, Jeannin P
Blood, 2007-09-11;110(13):4319-30.
Species: Human
Sample Types: Cell Culture Supernates
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Respiratory syncytial virus synergizes with Th2 cytokines to induce optimal levels of TARC/CCL17.
Authors: Monick MM, Powers LS, Hassan I, Groskreutz D, Yarovinsky TO, Barrett CW, Castilow EM, Tifrea D, Varga SM, Hunninghake GW
J. Immunol., 2007-08-01;179(3):1648-58.
Species: Human
Sample Types: Cell Culture Supernates
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Airway remodelling in children with cystic fibrosis.
Authors: Hilliard TN, Regamey N, Shute JK, Nicholson AG, Alton EW, Bush A, Davies JC
Thorax, 2007-05-25;62(12):1074-80.
Species: Human
Sample Types: BALF
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Mast cells regulate procollagen I (alpha 1) production by bronchial fibroblasts derived from subjects with asthma through IL-4/IL-4 delta 2 ratio.
Authors: Plante S, Semlali A, Joubert P, Bissonnette E, Laviolette M, Hamid Q, Chakir J
J. Allergy Clin. Immunol., 2006-02-21;117(6):1321-7.
Species: Human
Sample Types: Cell Culture Supernates
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A depressed peripheral cellular immune response is related to symptomatic neurocysticercosis.
Authors: Chavarria A, Fleury A, Bobes RJ, Morales J, Fragoso G, Sciutto E
Microbes Infect., 2006-01-19;8(4):1082-9.
Species: Human
Sample Types: Cell Culture Supernates
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Association of type 2 cytokines with hepatic fibrosis in human Schistosoma mansoni infection.
Authors: de Jesus AR, Magalhaes A, Miranda DG, Miranda RG, Araujo MI, de Jesus AA, Silva A, Santana LB, Pearce E, Carvalho EM
Infect. Immun., 2004-06-01;72(6):3391-7.
Species: Human
Sample Types: Cell Culture Supernates
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easy to use, IL-13 detectable in house dust mite stimulated PBMCs.
