Mouse CCL11/Eotaxin 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 CCL11/Eotaxin. 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: CCL11/Eotaxin
CCL11/Eotaxin is a chemokine that interacts with CCR3 to induce eosinophil, basophil, dendritic cell, and Th2 cell chemoattraction during inflammatory and allergic reactions. Its production is upregulated in airway epithelium, alveolar macrophages, eosinophils, smooth muscle cells, chondrocytes, and fibroblasts following allergen or inflammatory challenge. CCL11 is also involved in the growth of myeloid cell progenitors and the differentiation of mast cells.
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 CCL11/Eotaxin 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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Pro-cancerogenic effects of spontaneous and drug-induced senescence of ovarian cancer cells in vitro and in vivo: a comparative analysis
Authors: S Rutecki, P Szulc, M Paku?a, P Uruski, A Radziemski, E Naumowicz, R Moszy?ski, A Tykarski, J Miku?a-Pie, K Ksi??ek
Journal of ovarian research, 2022-07-26;15(1):87.
Species: Human
Sample Types: Cell Culture Supernates
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Exogenous leptin enhances markers of airway fibrosis in a mouse model of chronic allergic airways disease
Authors: MD Ihrie, VL McQuade, JT Womble, A Hegde, MS McCravy, CVG Lacuesta, RM Tighe, LG Que, JKL Walker, JL Ingram
Respiratory Research, 2022-05-24;23(1):131.
Species: Mouse
Sample Types: BALF
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DRP1-Mediated Mitochondrial Fission Regulates Lung Epithelial Response to Allergen
Authors: SR Bruno, A Kumar, ZF Mark, R Chandrasek, E Nakada, N Chamberlai, B Mihavics, J Walzer, J Cahoon, AE Dixon, B Cunniff, V Anathy
International Journal of Molecular Sciences, 2021-10-15;22(20):.
Species: Mouse
Sample Types: Tissue Homeogenates
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Ablation of RhoA impairs Th17 cell differentiation and alleviates house dust mite-triggered allergic airway inflammation
Authors: JQ Yang, KW Kalim, Y Li, Y Zheng, F Guo
J. Leukoc. Biol., 2019-07-01;0(0):.
Species: Mouse
Sample Types: BALF
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Conjugated bile acids attenuate allergen-induced airway inflammation and hyperresposiveness by inhibiting UPR transducers
Authors: EM Nakada, NR Bhakta, BR Korwin-Mih, A Kumar, N Chamberlai, SR Bruno, DG Chapman, SM Hoffman, N Daphtary, M Aliyeva, CG Irvin, AE Dixon, PG Woodruff, S Amin, ME Poynter, DH Desai, V Anathy
JCI Insight, 2019-05-02;4(9):.
Species: Mouse
Sample Types: Tissue Homogenates
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Inhalation of the prodrug PI3K inhibitor CL27c improves lung function in asthma and fibrosis
Authors: CC Campa, RL Silva, JP Margaria, T Pirali, MS Mattos, LR Kraemer, DC Reis, G Grosa, F Copperi, EM Dalmarco, RCP Lima-Júnio, S Aprile, V Sala, F Dal Bello, DS Prado, JC Alves-Filh, C Medana, GD Cassali, GC Tron, MM Teixeira, E Ciraolo, RC Russo, E Hirsch
Nat Commun, 2018-12-12;9(1):5232.
Species: Mouse
Sample Types: Tissue Homogenates
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Repeated Allergen Exposure in A/J Mice Causes Steroid-Insensitive Asthma via a Defect in Glucocorticoid Receptor Bioavailability
Authors: MF Serra, AC Cotias, CRR Pão, JB Daleprane, PB Jurgilas, GC Couto, EA Anjos-Valo, RSB Cordeiro, VF Carvalho, PMR Silva, MA Martins
J. Immunol., 2018-06-18;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
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Probiotic treatment during neonatal age provides optimal protection against experimental asthma through the modulation of microbiota and T cells: Neonatal exposure to probiotics prevents asthma
Authors: CFCG Nunes, JS Nogueira, FB Canto, PHO Vianna, BT Ciambarell, PMRE Silva, KR Miranda, LA Lobo, RMCP Domingues, M Busch, GC Atella, AM Vale, M Bellio, A Nóbrega, R Fucs
Int. Immunol., 2018-04-03;0(0):.
Species: Mouse
Sample Types: BALF
Applications: ELISA Capture -
Schistosoma japonicum infection downregulates house dust mite-induced allergic airway inflammation in mice
Authors: S Qiu, X Fan, Y Yang, P Dong, W Zhou, Y Xu, Y Zhou, F Guo, Y Zheng, JQ Yang
PLoS ONE, 2017-06-14;12(6):e0179565.
Species: Mouse
Sample Types: BALF
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Regulatory effects of Interleukin (IL)-15 on allergen-induced airway obstruction
Authors: SU Venkatesha, X Zhu, P Rajavelu, R Niranjan, M Manohar, AK Verma, JA Lasky, A Mishra
J. Allergy Clin. Immunol., 2017-06-09;0(0):.
Species: Mouse
Sample Types: BALF
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Factor XI deficiency enhances the pulmonary allergic response to house dust mite in mice independent of factor XII
Authors: Ingrid Stroo
Am. J. Physiol. Lung Cell Mol. Physiol, 2016-12-02;0(0):ajplung.00320.
Species: Mouse
Sample Types: Tissue Homogenates
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Regulation of eosinophilia and allergic airway inflammation by the glycan-binding protein galectin-1
Proc Natl Acad Sci USA, 2016-07-25;0(0):.
Species: Mouse
Sample Types: BALF
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Paired Ig-like Receptor B Inhibits IL-13-Driven Eosinophil Accumulation and Activation in the Esophagus
Authors: Netali Ben Baruch
J Immunol, 2016-06-20;0(0):.
Species: Mouse
Sample Types: BALF
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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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Mice deficient in the St3gal3 gene product alpha2,3 sialyltransferase (ST3Gal-III) exhibit enhanced allergic eosinophilic airway inflammation.
Authors: Kiwamoto T, Brummet M, Wu F, Motari M, Smith D, Schnaar R, Zhu Z, Bochner B
J Allergy Clin Immunol, 2013-07-02;133(1):240-7.e1-3.
Species: Mouse
Sample Types: BALF
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Intestinal CCL11 and eosinophilic inflammation is regulated by myeloid cell-specific RelA/p65 in mice.
Authors: Waddell A, Ahrens R, Tsai Y, Sherrill J, Denson L, Steinbrecher K, Hogan S
J Immunol, 2013-04-05;190(9):4773-85.
Species: Mouse
Sample Types: Cell Culture Supernates
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Control of allergen-induced inflammation and hyperresponsiveness by the metalloproteinase ADAMTS-12.
Authors: Paulissen G, El Hour M, Rocks N, Gueders M, Bureau F, Foidart J, Lopez-Otin C, Noel A, Cataldo D
J Immunol, 2012-09-07;189(8):4135-43.
Species: Mouse
Sample Types: Tissue Homogenates
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Role of Toll interleukin-1 receptor (IL-1R) 8, a negative regulator of IL-1R/Toll-like receptor signaling, in resistance to acute Pseudomonas aeruginosa lung infection.
Authors: Veliz Rodriguez T, Moalli F, Polentarutti N, Paroni M, Bonavita E, Anselmo A, Nebuloni M, Mantero S, Jaillon S, Bragonzi A, Mantovani A, Riva F, Garlanda C
Infect. Immun., 2011-10-24;80(1):100-9.
Species: Mouse
Sample Types: Serum
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Alpha-Galactosylceramide-induced airway eosinophilia is mediated through the activation of NKT cells.
Authors: Chuang YH, Wang TC, Jen HY, Yu AL, Chiang BL
J. Immunol., 2011-03-07;186(8):4687-92.
Species: Mouse
Sample Types: BALF
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A natural protective function of invariant NKT cells in a mouse model of innate-cell-driven lung inflammation.
Authors: Bourgeois EA, Levescot A, Diem S, Chauvineau A, Berges H, Milpied P, Lehuen A, Damotte D, Gombert JM, Schneider E, Girard JP, Gourdy P, Herbelin A
Eur. J. Immunol., 2011-01-11;41(2):299-305.
Species: Mouse
Sample Types: BALF
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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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Impact of adiponectin deficiency on pulmonary responses to acute ozone exposure in mice.
Authors: Zhu M, Hug C, Kasahara DI
Am. J. Respir. Cell Mol. Biol., 2009-11-13;43(4):487-97.
Species: Mouse
Sample Types: BALF
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IL-6 is required for airway mucus production induced by inhaled fungal allergens.
Authors: Neveu WA, Allard JB, Dienz O, Wargo MJ, Ciliberto G, Whittaker LA, Rincon M
J. Immunol., 2009-07-10;183(3):1732-8.
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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Coexposure to environmental tobacco smoke increases levels of allergen-induced airway remodeling in mice.
Authors: Min MG, Song DJ, Miller M, Cho JY, McElwain S, Ferguson P, Broide DH
J. Immunol., 2007-04-15;178(8):5321-8.
Species: Mouse
Sample Types: BALF
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Toll-like receptor-2-mediated C-C chemokine receptor 3 and eotaxin-driven eosinophil influx induced by Mycobacterium bovis BCG pleurisy.
Authors: D'Avila H, Almeida PE, Roque NR, Castro-Faria-Neto HC, Bozza PT
Infect. Immun., 2006-12-11;75(3):1507-11.
Species: Mouse
Sample Types: Pleural Effusion
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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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Induced trefoil factor family 1 expression by trans-differentiating Clara cells in a murine asthma model.
Authors: Kouznetsova I, Chwieralski CE, Balder R, Hinz M, Braun A, Krug N, Hoffmann W
Am. J. Respir. Cell Mol. Biol., 2006-09-21;36(3):286-95.
Species: Mouse
Sample Types: BALF
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Increased pulmonary responses to acute ozone exposure in obese db/db mice.
Authors: Lu FL, Johnston RA, Flynt L, Theman TA, Terry RD, Schwartzman IN, Lee A, Shore SA
Am. J. Physiol. Lung Cell Mol. Physiol., 2005-12-22;290(5):L856-65.
Species: Mouse
Sample Types: Serum
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