Mouse IL-17E 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 IL-17E. 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-17E/IL-25
The IL-17 family is comprised of at least six proinflammatory cytokines that share a conserved cysteine-knot structure but diverge at the N-terminus. IL-17 family members are glycoproteins secreted as dimers that induce local cytokine production and recruit granulocytes to sites of inflammation. IL-17 is induced by IL-15 and IL-23, mainly in activated CD4+ T cells distinct from Th1 or Th2 cells. IL-17F is the most homologous to IL-17, but is induced only by IL-23 in activated monocytes. IL-17B binds the IL-17B receptor, but not the IL-17 receptor; it is most homologous with IL-17D, which is expressed by resting CD4+ T cells and CD19+ B cells. IL-17E is mainly produced by Th2 cells and recruits eosinophils to lung tissue. IL-17C has a very restricted expression pattern but has been detected in adult prostate and fetal kidney libraries.
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 IL-17E 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.
25
Citations: Showing 1 - 10
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Microbiota encoded fatty-acid metabolism expands tuft cells to protect tissues homeostasis during Clostridioides difficile infection in the large intestine
Authors: Kellogg, TD;Ceglia, S;Mortzfeld, BM;Zeamer, AL;Foley, SE;Ward, DV;Bhattarai, SK;McCormick, BA;Reboldi, A;Bucci, V;
bioRxiv : the preprint server for biology
Species: Mouse
Sample Types: Tissue Homogenates
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Increased hepatic interleukin-1, arachidonic acid, and reactive oxygen species mediate the protective potential of peptides shared by gut cysteine peptidases against Schistosoma mansoni infection in mice
Authors: H Tallima, R El Ridi
PloS Neglected Tropical Diseases, 2023-03-15;17(3):e0011164.
Species: Mouse
Sample Types: Tissue Homogenates
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Crosstalk between ILC2s and Th2 cells varies among mouse models
Authors: RK Gurram, D Wei, Q Yu, MJ Butcher, X Chen, K Cui, G Hu, M Zheng, X Zhu, J Oh, B Sun, JF Urban, K Zhao, WJ Leonard, J Zhu
Cell Reports, 2023-02-02;42(2):112073.
Species: Mouse
Sample Types: BALF
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Inhaled delivery of recombinant interferon-lambda restores allergic inflammation after development of asthma by controlling Th2- and Th17-cell-mediated immune responses
Authors: J Won, A Jo, CH Gil, S Kim, H Shin, H Jik Kim
International immunopharmacology, 2022-08-27;112(0):109180.
Species: Mouse
Sample Types: BALF
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Macrophage-intrinsic DUOX1 contributes to type 2 inflammation and mucus metaplasia during allergic airway disease
Authors: CR Morris, A Habibovic, CM Dustin, C Schiffers, MC Lin, JL Ather, YMW Janssen-He, ME Poynter, O Utermohlen, M Krönke, A van der Vl
Mucosal Immunology, 2022-06-02;0(0):.
Species: Mouse
Sample Types: BALF
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Eosinophil extracellular traps drive asthma progression through neuro-immune signals
Authors: Y Lu, Y Huang, J Li, J Huang, L Zhang, J Feng, J Li, Q Xia, Q Zhao, L Huang, S Jiang, S Su
Nature Cell Biology, 2021-10-06;23(10):1060-1072.
Species: Mouse
Sample Types: BALF
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Diesel exhaust particles increase nasal symptoms and IL-17A in house dust mite-induced allergic mice
Authors: HJ Jung, YK Ko, WS Shim, HJ Kim, DY Kim, CS Rhee, MK Park, DH Han
Scientific Reports, 2021-08-11;11(1):16300.
Species: Mouse
Sample Types: Cell Culture Supernates
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Tumor suppressor p53 regulates intestinal type 2 immunity
Authors: CY Chang, J Wang, Y Zhao, J Liu, X Yang, X Yue, H Wang, F Zhou, JM Inclan-Ric, JJ Ponessa, P Xie, L Zhang, MC Siracusa, Z Feng, W Hu
Nature Communications, 2021-06-07;12(1):3371.
Species: Mouse
Sample Types: Tissue Homogenates
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Interleukin (IL)-33 is dispensable for Schistosoma mansoni worm maturation and the maintenance of egg-induced pathology in intestines of infected mice
Authors: JPK Mukendi, R Nakamura, S Uematsu, S Hamano
Parasites & vectors, 2021-01-22;14(1):70.
Species: Mouse
Sample Types: Tissue Homogenates
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Role of biomarkers and effect of FIP-fve in acute and chronic animal asthma models
Authors: CT Wu, YT Lee, MS Ku, KH Lue
J Microbiol Immunol Infect, 2020-07-28;0(0):.
Species: Mouse
Sample Types: BALF
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Exophilin-5 regulates allergic airway inflammation by controlling IL-33-mediated Th2 responses
Authors: K Okunishi, H Wang, M Suzukawa, R Ishizaki, E Kobayashi, M Kihara, T Abe, JI Miyazaki, M Horie, A Saito, H Saito, S Nakae, T Izumi
J. Clin. Invest., 2020-07-01;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
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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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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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Evaluation of murine lung epithelial cells (TC-1 JHU-1) line to develop Th2-promoting cytokines IL-25/IL-33/TSLP and genes Tlr2/Tlr4 in response to Aspergillus fumigatus
Authors: AR Khosravi, H Shokri, S Hassan Al-, F Ghafarifar
J Mycol Med, 2018-03-07;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Dysregulated invertebrate tropomyosin-dectin-1 interaction confers susceptibility to allergic diseases
Authors: N Gour, S Lajoie, U Smole, M White, D Hu, P Goddard, S Huntsman, C Eng, A Mak, S Oh, JH Kim, A Sharma, S Plante, IH Salem, Y Resch, X Xiao, N Yao, A Singh, S Vrtala, J Chakir, EG Burchard, AP Lane, M Wills-Karp
Sci Immunol, 2018-02-23;3(20):.
Species: Mouse
Sample Types: BALF
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Microbiome-mediated neutrophil recruitment via CXCR2 and protection from amebic colitis
Authors: K Watanabe, CA Gilchrist, MJ Uddin, SL Burgess, MM Abhyankar, SN Moonah, Z Noor, JR Donowitz, BN Schneider, T Arju, E Ahmed, M Kabir, M Alam, R Haque, P Pramoonjag, B Mehrad, WA Petri
PLoS Pathog., 2017-08-17;13(8):e1006513.
Species: Mouse
Sample Types: Tissue Homogenates
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STAT1 Represses Cytokine-Producing Group 2 and Group 3 Innate Lymphoid Cells during Viral Infection
Authors: MT Stier, K Goleniewsk, JY Cephus, DC Newcomb, TP Sherrill, KL Boyd, MH Bloodworth, ML Moore, K Chen, JK Kolls, RS Peebles
J. Immunol., 2017-06-02;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
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Intelectin contributes to allergen-induced IL-25, IL-33, and TSLP expression and type 2 response in asthma and atopic dermatitis
Authors: L Yi, D Cheng, K Zhang, X Huo, Y Mo, H Shi, H Di, Y Zou, H Zhang, J Zhao, Y Xu, DJ Erle, G Zhen
Mucosal Immunol, 2017-02-22;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
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Interleukin-33 from Monocytes Recruited to the Lung Contributes to House Dust Mite-Induced Airway Inflammation in a Mouse Model
Authors: Hiroki Tashiro
PLoS ONE, 2016-06-16;11(6):e0157571.
Species: Mouse
Sample Types: Tissue Homogenates
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IL-22 Restrains Tapeworm-Mediated Protection against Experimental Colitis via Regulation of IL-25 Expression
Authors: JL Reyes, MR Fernando, F Lopes, G Leung, NL Mancini, CE Matisz, A Wang, DM McKay
PLoS Pathog, 2016-04-07;12(4):e1005481.
Species: Mouse
Sample Types: Cell Culture Supernates
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Lung-homing of endothelial progenitor cells and airway vascularization is only partially dependant on eosinophils in a house dust mite-exposed mouse model of allergic asthma.
Authors: Sivapalan, Nirooya, Wattie, Jennifer, Inman, Mark D, Sehmi, Roma
PLoS ONE, 2014-10-03;9(10):e109991.
Species: Mouse
Sample Types: Tissue Homogenates
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The Toll-like receptor 5 ligand flagellin promotes asthma by priming allergic responses to indoor allergens.
Authors: Wilson, Rhonda H, Maruoka, Shuichir, Whitehead, Gregory, Foley, Julie F, Flake, Gordon P, Sever, Michelle, Zeldin, Darryl C, Kraft, Monica, Garantziotis, Stavros, Nakano, Hideki, Cook, Donald N
Nat Med, 2012-10-14;18(11):1705-10.
Species: Mouse
Sample Types: BALF
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Epithelial cell-derived IL-25, but not Th17 cell-derived IL-17 or IL-17F, Is crucial for murine asthma.
Authors: Suzukawa M, Morita H, Nambu A, Arae K, Shimura E, Shibui A, Yamaguchi S, Suzukawa K, Nakanishi W, Oboki K, Kajiwara N, Ohno T, Ishii A, Korner H, Cua D, Suto H, Yoshimoto T, Iwakura Y, Yamasoba T, Ohta K, Sudo K, Saito H, Okumura K, Broide D, Matsumoto K, Nakae S
J Immunol, 2012-08-31;189(7):3641-52.
Species: Mouse
Sample Types: BALF
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Interleukin-1alpha controls allergic sensitization to inhaled house dust mite via the epithelial release of GM-CSF and IL-33.
J. Exp. Med., 2012-07-16;209(8):1505-17.
Species: Mouse
Sample Types: Cell Culture Supernates
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IL-33-responsive lineage- CD25+ CD44(hi) lymphoid cells mediate innate type 2 immunity and allergic inflammation in the lungs.
Authors: Bartemes KR, Iijima K, Kobayashi T, Kephart GM, McKenzie AN, Kita H
J. Immunol., 2011-12-23;188(3):1503-13.
Species: Mouse
Sample Types: BALF
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First time using this kit which has been in the lab for a while. Excellent results. Standards come up very nicely. Instructions clear and easy to follow. 5 stars from me.
