Human IL-5 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-5
IL-5 (Interleukin-5) is a secreted cytokine that is expressed as a covalent dimer. It is produced by activated Th2 cells, eosinophils, mast cells, and iNKT cells and promotes eosinophilic and basophilic differentiation and activation. IL-5 signals through a receptor complex containing IL-5 R alpha and the Common beta Chain/CD131 on eosinophils and basophils. Naturally occurring soluble forms of IL-5 R alpha retain the ability to bind IL-5 and limit its bioavailability.
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-5 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.
26
Citations: Showing 1 - 10
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Neutrophil extracellular traps promote immunopathogenesis of virus-induced COPD exacerbations
Authors: Katsoulis, O;Toussaint, M;Jackson, MM;Mallia, P;Footitt, J;Mincham, KT;Meyer, GFM;Kebadze, T;Gilmour, A;Long, M;Aswani, AD;Snelgrove, RJ;Johnston, SL;Chalmers, JD;Singanayagam, A;
Nature communications
Species: Human
Sample Types: Sputum
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Cognitive function is associated with performance in time up and go test and with leptin blood levels in community-dwelling older women
Authors: da Costa Teixeira, LA;Soares, LA;Lima, LP;Avelar, NCP;de Moura, JA;Leopoldino, AAO;Figueiredo, PHS;Parentoni, AN;Mendonça, VA;Lacerda, ACR;
Scientific reports
Species: Human
Sample Types: Plasma
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The strong inverse association between plasma concentrations of soluble tumor necrosis factor receptors type 1 with adiponectin/leptin ratio in older women
Authors: Augusto da Costa Teixeira, L;Rocha-Vieira, E;Aparecida Soares, L;Mota de Oliveira, F;Aparecida Oliveira Leopoldino, A;Netto Parentoni, A;Amaral Mendonça, V;Cristina Rodrigues Lacerda, A;
Cytokine
Species: Human
Sample Types: Plasma
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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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Macrophages derived from LPS-stimulated monocytes from individuals with subclinical atherosclerosis were characterized by increased pro-inflammatory activity
Authors: Nikiforov, NG;Kirichenko, TV;Kubekina, MV;Chegodaev, YS;Zhuravlev, AD;Ilchuk, LA;Nikolaeva, MA;Arefieva, AS;Popov, MA;Verkhova, SS;Bagheri Ekta, M;Orekhov, AN;
Cytokine
Species: Human
Sample Types: Cell Culture Supernates
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Inflammatory biomarkers at different stages of Sarcopenia in older women
Authors: da Costa Teixeira, LA;Avelar, NCP;Peixoto, MFD;Parentoni, AN;Santos, JMD;Pereira, FSM;Danielewicz, AL;Leopoldino, AAO;Costa, SP;Arrieiro, AN;Soares, LA;da Silva Lage, VK;Prates, ACN;Taiar, R;de Carvalho Bastone, A;Oliveira, VC;Oliveira, MX;Costa, HS;Nobre, JNP;Brant, FP;Duarte, TC;Figueiredo, PHS;Mendonça, VA;Lacerda, ACR;
Scientific reports
Species: Human
Sample Types: Plasma
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Adiponectin Is a Contributing Factor of Low Appendicular Lean Mass in Older Community-Dwelling Women: A Cross-Sectional Study
Authors: LAC Teixeira, JM Dos Santos, AN Parentoni, LP Lima, TC Duarte, FP Brant, CDC Neves, FSM Pereira, NCP Avelar, AL Danielewic, AAO Leopoldino, SP Costa, AN Arrieiro, LA Soares, ACN Prates, JNP Nobre, A de Carvalh, VC de Oliveir, MX Oliveira, PH Scheidt Fi, HS Costa, V Amaral Men, R Taiar, AC Rodrigues
Journal of Clinical Medicine, 2022-12-02;11(23):.
Species: Human
Sample Types: Plasma
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Substrate stiffness engineered to replicate disease conditions influence senescence and fibrotic responses in primary lung fibroblasts
Authors: KEC Blokland, M Nizamoglu, H Habibie, T Borghuis, M Schuliga, BN Melgert, DA Knight, CA Brandsma, SD Pouwels, JK Burgess
Frontiers in Pharmacology, 2022-11-03;13(0):989169.
Species: Human
Sample Types: Cell Culture Supernates
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Anti-inflammatory effects of an autologous gold-based serum therapy in osteoarthritis patients
Authors: J Feldt, AJ Donaubauer, J Welss, U Schneider, US Gaipl, F Paulsen
Scientific Reports, 2022-03-03;12(1):3560.
Species: Human
Sample Types: Serum
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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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Effectiveness and safety of mepolizumab in combination with corticosteroids in patients with eosinophilic granulomatosis with polyangiitis
Authors: M Ueno, I Miyagawa, K Nakano, S Iwata, K Hanami, S Fukuyo, S Kubo, Y Miyazaki, A Kawabe, H Yoshinari, S Nakayamada, Y Tanaka
Arthritis Research & Therapy, 2021-03-16;23(1):86.
Species: Human
Sample Types: Serum
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New Biomarker in Chagas Disease: Extracellular Vesicles Isolated from Peripheral Blood in Chronic Chagas Disease Patients Modulate the Human Immune Response
Authors: RP Madeira, LM Dal'Mas Ro, P de Cássia, C Mady, BM Ianni, AC Torrecilha
Journal of Immunology Research, 2021-01-11;2021(0):6650670.
Species: Human
Sample Types: Cell Culture Supernates
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Altered plasma levels of &betaC and &gammaC chain cytokines and post-treatment modulation in tuberculous lymphadenitis
Authors: GR Kathamuthu, K Moideen, R Sridhar, D Baskaran, S Babu
Cytokine, 2020-12-17;138(0):155405.
Species: Human
Sample Types: Plasma
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Salivary IL-8 and IL-1RA changes induced by exercise in cold-dry and normal conditions
Authors: I Patlan, MP Jeffrey, D Lewis, J Green-John, S Dogra
Cytokine, 2020-10-28;137(0):155347.
Species: Human
Sample Types: Saliva
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Altered miR-155 expression in Allergic Asthmatic airways
Authors: C Malmhäll, K Johansson, C Winkler, S Alawieh, L Ekerljung, M Rådinger
Scand. J. Immunol, 2017-04-01;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Genetic, immunological, and clinical features of patients with bacterial and fungal infections due to inherited IL-17RA deficiency
Proc. Natl. Acad. Sci. U.S.A, 2016-12-07;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Human CD4+ T cell responses to the dog major allergen Can f 1 and its human homologue tear lipocalin resemble each other.
Authors: Liukko A, Kinnunen T, Rytkonen-Nissinen M, Kailaanmaki A, Randell J, Maillere B, Virtanen T
PLoS ONE, 2014-05-29;9(5):e98461.
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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An ACT1 mutation selectively abolishes interleukin-17 responses in humans with chronic mucocutaneous candidiasis.
Authors: Boisson B, Wang C, Pedergnana V, Wu L, Cypowyj S, Rybojad M, Belkadi A, Picard C, Abel L, Fieschi C, Puel A, Li X, Casanova J
Immunity, 2013-10-10;39(4):676-86.
Species: Human
Sample Types: Cell Culture Supernates
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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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Natural killer cell-mediated cytokine response among HIV-positive south Indians with pulmonary tuberculosis.
Authors: Ramana Rao PV, Rajasekaran S, Raja A
J. Interferon Cytokine Res., 2010-01-01;30(1):33-42.
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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Neuroimmune crosstalk in asthma: dual role of the neurotrophin receptor p75NTR.
Authors: Nassenstein C, Kammertoens T, Veres TZ, Uckert W, Spies E, Fuchs B, Krug N, Braun A
J. Allergy Clin. Immunol., 2007-08-22;120(5):1089-96.
Species: Mouse
Sample Types: BALF
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An investigation of auto-reactivity after head injury.
Authors: Cox AL, Coles AJ, Nortje J, Bradley PG, Chatfield DA, Thompson SJ, Menon DK
J. Neuroimmunol., 2006-03-06;174(1):180-6.
Species: Human
Sample Types: Cell Culture Supernates
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Synthesis and biological evaluation of 1,2,4-triazinylphenylalkylthiazolecarboxylic acid esters as cytokine-inhibiting antedrugs with strong bronchodilating effects in an animal model of asthma.
Authors: Freyne EJ, Lacrampe JF, Deroose F, Boeckx GM, Willems M, Embrechts W, Coesemans E, Willems JJ, Fortin JM, Ligney Y, Dillen LL, Cools WF, Goossens J, Corens D, De Groot A, Van Wauwe JP
J. Med. Chem., 2005-03-24;48(6):2167-75.
Species: Human
Sample Types: Cell Culture Supernates
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easy to use kit. IL-5 detectable in PBMCs treated with house dust mite.