Mouse CCL17/TARC 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 mouse CCL17/TARC. 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: CCL17/TARC
CCL17/TARC is a chemokine that interacts with CCR4 and CCR8 to induce the chemoattraction of activated Th2 cells, basophils, and NK cells. It is produced by thymic dendritic cells, lymph node dendritic cells, monocytes, CD4+ T cells, keratinocytes, fibroblasts, bronchial epithelial cells, and Reed-Sternberg cells. CCL17 promotes Th2 cell recruitment to sites of inflammation and induces platelet aggregation and degranulation.
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 Mouse CCL17/TARC 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.
16
Citations: Showing 1 - 10
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Co-Application with Tannic Acid Prevents Transdermal Sensitization to Ovalbumin in Mice
Authors: E Izumi, N Tanahashi, S Kinugasa, S Hidaka, N Zaima, T Moriyama
International Journal of Molecular Sciences, 2022-04-01;23(7):.
Species: Mouse
Sample Types: Tissue Homogenates
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EBV+ tumors exploit tumor cell-intrinsic and -extrinsic mechanisms to produce regulatory T cell-recruiting chemokines CCL17 and CCL22
Authors: A Jorapur, LA Marshall, S Jacobson, M Xu, S Marubayash, M Zibinsky, DX Hu, O Robles, JJ Jackson, V Baloche, P Busson, D Wustrow, DG Brockstedt, O Talay, PD Kassner, G Cutler
PloS Pathogens, 2022-01-13;18(1):e1010200.
Species: Mouse
Sample Types: Cell Culture Supernates
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Dietary galacto-oligosaccharides prevent airway eosinophilia and hyperresponsiveness in a murine house dust mite-induced asthma model.
Authors: Verheijden K, Willemsen L, Braber S, Leusink-Muis T, Delsing D, Garssen J, Kraneveld A, Folkerts G
Respir Res, 2015-02-07;16(0):17.
Species: Mouse
Sample Types: Tissue Homogenates
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Protease-activated receptor-2 activation contributes to house dust mite-induced IgE responses in mice.
Authors: Post, Sijranke, Heijink, Irene H, Petersen, Arjen H, de Bruin, Harold G, van Oosterhout, Antoon J, Nawijn, Martijn
PLoS ONE, 2014-03-20;9(3):e91206.
Species: Mouse
Sample Types: Tissue Homogenates
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MicroRNA-155 is essential for T(H)2-mediated allergen-induced eosinophilic inflammation in the lung.
Authors: Malmhall C, Alawieh S, Lu Y, Sjostrand M, Bossios A, Eldh M, Radinger M
J Allergy Clin Immunol, 2013-12-24;133(5):1429-38, 1438.
Species: Mouse
Sample Types: Tissue Homogenates
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Influence of hypoxia-inducible factor 1-alpha on dendritic cell differentiation and migration.
Authors: Kohler T, Reizis B, Johnson RS, Weighardt H, Forster I
Eur. J. Immunol., 2012-05-01;42(5):1226-36.
Species: Mouse
Sample Types: Cell Culture Supernates
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CCL17 controls mast cells for the defense against filarial larval entry.
Authors: Specht S, Frank JK, Alferink J, Dubben B, Layland LE, Denece G, Bain O, Forster I, Kirschning CJ, Martin C, Hoerauf A
J. Immunol., 2011-03-11;186(8):4845-52.
Species: Mouse
Sample Types: Thoracic Cavity Fluid
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Inflammatory chemokine release of astrocytes in vitro is reduced by all-trans retinoic acid.
Authors: van Neerven S, Regen T, Wolf D, Nemes A, Johann S, Beyer C, Hanisch UK, Mey J
J. Neurochem., 2010-06-16;114(5):1511-26.
Species: Mouse
Sample Types: Cell Culture Supernates
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Human T-lymphotropic virus type 1-induced CC chemokine ligand 22 maintains a high frequency of functional FoxP3+ regulatory T cells.
Authors: Toulza F, Nosaka K, Tanaka Y
J. Immunol., 2010-06-04;185(1):183-9.
Species: Human
Sample Types: Cell Culture Supernates
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IL-12 produced by dendritic cells augments CD8+ T cell activation through the production of the chemokines CCL1 and CCL17.
Authors: Henry CJ, Ornelles DA, Mitchell LM, Brzoza-Lewis KL, Hiltbold EM
J. Immunol., 2008-12-15;181(12):8576-84.
Species: Mouse
Sample Types: Cell Culture Supernates
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Distinct roles for IL-13 and IL-4 via IL-13 receptor alpha1 and the type II IL-4 receptor in asthma pathogenesis.
Authors: Munitz A, Brandt EB, Mingler M, Finkelman FD, Rothenberg ME
Proc. Natl. Acad. Sci. U.S.A., 2008-05-14;105(20):7240-5.
Species: Mouse
Sample Types: BALF
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Surfactant protein D alters allergic lung responses in mice and human subjects.
Authors: Brandt EB, Mingler MK, Stevenson MD, Wang N, Khurana Hershey GK, Whitsett JA, Rothenberg ME
J. Allergy Clin. Immunol., 2008-03-19;121(5):1140-1147.e2.
Species: Mouse
Sample Types: BALF
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Adenovirus IL-13-induced airway disease in mice: a corticosteroid-resistant model of severe asthma.
Authors: Therien AG, Bernier V, Weicker S, Tawa P, Falgueyret JP, Mathieu MC, Honsberger J, Pomerleau V, Robichaud A, Stocco R, Dufresne L, Houshyar H, Lafleur J, Ramachandran C, O'Neill GP, Slipetz D, Tan CM
Am. J. Respir. Cell Mol. Biol., 2008-02-07;39(1):26-35.
Species: Mouse
Sample Types: BALF
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IL-1R1/MyD88 signaling and the inflammasome are essential in pulmonary inflammation and fibrosis in mice.
Authors: Gasse P, Mary C, Guenon I, Noulin N, Charron S, Schnyder-Candrian S, Schnyder B, Akira S, Quesniaux VF, Lagente V, Ryffel B, Couillin I
J. Clin. Invest., 2007-12-01;117(12):3786-99.
Species: Mouse
Sample Types: BALF
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Interleukin-17 is a negative regulator of established allergic asthma.
Authors: Schnyder-Candrian S, Togbe D, Couillin I, Mercier I, Brombacher F, Quesniaux V, Fossiez F, Ryffel B, Schnyder B
J. Exp. Med., 2006-11-13;203(12):2715-25.
Species: Mouse
Sample Types: Cell Culture Supernates
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Immunosuppressive effects of CCL17 on pulmonary antifungal responses during pulmonary invasive aspergillosis.
Authors: Carpenter KJ, Hogaboam CM
Infect. Immun., 2005-11-01;73(11):7198-207.
Species: Mouse
Sample Types: Tissue Homogenates
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CCL17 induced by IL-13 recombinant protein in mouse airway