Mouse IL-22 DuoSet ELISA
Mouse IL-22 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-22. 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 3 (5 plates): (Catalog # DY009) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 3.
Normal Goat Serum: (Catalog # DY005)
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 # DY004), or 5% Tween 20 in PBS, 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)
Normal Goat Serum: (Catalog # DY005)
Scientific Data
Product Datasheets
Preparation and Storage
Background: IL-22
IL-22 (Interleukin-22) is a cytokine that induces the production of reactive oxygen species, IL-6, IL-10, and TNF-alpha as well as neutrophil infiltration during inflammation. It also supports the integrity of epithelial barriers and induces epithelial cell proliferation during wound healing. IL-22 signals through a receptor complex consisting of IL-22 R and IL-10 R beta. IL-10 R beta is a shared component of the receptor complexes for IL-10, IL-26, IL-28, and IL-29. IL-22 additionally binds to IL-22BP which blocks the interaction of IL-22 with IL-22 R.
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 of 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 with NGS, 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-22 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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ERdj5 protects goblet cells from endoplasmic reticulum stress-mediated apoptosis under inflammatory conditions
Authors: H Jeong, EH Hong, JH Ahn, J Cho, JH Jeong, CW Kim, BI Yoon, JH Koo, YY Park, YM Yang, T Iwawaki, BA Vallance, SY Chang, HJ Ko
Experimental & Molecular Medicine, 2023-02-09;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
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Antimicrobial overproduction sustains intestinal inflammation by inhibiting Enterococcus colonization
Authors: KK Jang, T Heaney, M London, Y Ding, F Yeung, D Ercelen, YH Chen, J Axelrad, S Gurunathan, A Marijke Ke, ME Griffin, HC Hang, K Cadwell
bioRxiv : the preprint server for biology, 2023-02-01;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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IL-22 regulates inflammatory responses to agricultural dust-induced airway inflammation
Authors: A Ulu, S Sveiven, A Bilg, JV Velazquez, M Diaz, M Mukherjee, AG Yuil-Valde, S Kota, A Burr, A Najera, TM Nordgren
Toxicology and Applied Pharmacology, 2022-05-05;0(0):116044.
Species: Mouse
Sample Types: BALF
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ADP-ribosylating adjuvant reveals plasticity in cDC1 cells that drive mucosal Th17 cell development and protection against influenza virus infection
Authors: M Arabpour, C Lebrero-Fe, K Schön, A Strömberg, V Börjesson, K Lahl, M Ballegeer, X Saelens, D Angeletti, W Agace, N Lycke
Mucosal Immunology, 2022-04-13;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Effects of repeated infections with non-typeable Haemophilus influenzae on lung in vitamin D deficient and smoking mice
Authors: J Serré, AT Tanjeko, C Mathyssen, T Heigl, A Sacreas, DP Cook, E Verbeken, K Maes, J Verhaegen, C Pilette, J Vanoirbeek, C Gysemans, C Mathieu, B Vanaudenae, W Janssens, G Gayan-Rami
Respiratory Research, 2022-03-02;23(1):40.
Species: Mouse
Sample Types: Tissue Homogenates
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Geographic differences in gut microbiota composition impact susceptibility to enteric infection
Authors: AM Porras, Q Shi, H Zhou, R Callahan, G Montenegro, N Solomons, IL Brito
Cell Reports, 2021-07-27;36(4):109457.
Species: Mouse
Sample Types: Tissue Supernates
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Aryl hydrocarbon receptor (Ahr)-dependent Il-22 expression by type 3 innate lymphoid cells control of acute joint inflammation
Authors: R Nehmar, L Fauconnier, J Alves-Filh, D Togbe, A DeCauwer, S Bahram, M Le Bert, B Ryffel, P Georgel
Journal of Cellular and Molecular Medicine, 2021-03-18;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
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GM-CSF Calibrates Macrophage Defense and Wound Healing Programs during Intestinal Infection and Inflammation
Authors: T Castro-Dop, A Fleming, TW Dennison, JR Ferdinand, K Harcourt, BJ Stewart, Z Cader, ZK Tuong, C Jing, LSC Lok, RJ Mathews, A Portet, A Kaser, S Clare, MR Clatworthy
Cell Rep, 2020-07-07;32(1):107857.
Species: Mouse
Sample Types: Cell Culture Supernates
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Lactobacillus reuteri 5454 and Bifidobacterium animalis ssp. lactis 5764 improve colitis while differentially impacting dendritic cells maturation and antimicrobial responses
Authors: J Hrdý, J Alard, A Couturier-, O Boulard, D Boutillier, M Delacre, C Lapadatesc, A Cesaro, P Blanc, B Pot, B Ryffel, M Chamaillar, C Grangette
Sci Rep, 2020-03-24;10(1):5345.
Species: Mouse
Sample Types: Cell Culture Supernates
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PAD4-dependent NETs generation are indispensable for intestinal clearance of Citrobacter rodentium
Authors: P Saha, BS Yeoh, X Xiao, RM Golonka, V Singh, Y Wang, M Vijay-Kuma
Mucosal Immunol, 2019-02-01;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
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The Citrobacter rodentium type III secretion system effector EspO affects mucosal damage repair and antimicrobial responses
Authors: CN Berger, VF Crepin, TI Roumelioti, JC Wright, N Serafini, M Pevsner-Fi, L Yu, E Elinav, JP Di Santo, JS Choudhary, G Frankel
PLoS Pathog., 2018-10-26;14(10):e1007406.
Species: Mouse
Sample Types: Tissue Homogenates
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Integration of T Cell Receptor, Notch and Cytokine Signals Programs in Mouse ?? T Cell Effector Differentiation
Authors: P Zarin, TSH In, ELY Chen, J Singh, GW Wong, M Mohtashami, DL Wiest, MK Anderson, JC Zúñiga-Pfl
Immunol. Cell Biol., 2018-06-28;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Th17 cells are refractory to senescence and retain robust antitumor activity after long-term ex vivo expansion
Authors: JS Bowers, MH Nelson, K Majchrzak, SR Bailey, B Rohrer, AD Kaiser, C Atkinson, L Gattinoni, CM Paulos
JCI Insight, 2017-03-09;2(5):e90772.
Species: Mouse
Sample Types: Cell Culture Supernates
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T-bet overexpression regulates AHR-mediated Th-17 differentiation through an IFN?-independent pathway
Authors: Masahiro Yokosawa
Clin. Exp. Immunol, 2017-01-31;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Interleukin 7 immunotherapy improves host immunity and survival in a two-hit model of Pseudomonas aeruginosa pneumonia
J Leukoc Biol, 2016-09-14;0(0):.
Species: Mouse
Sample Types: Plasma
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Memory Th1 Cells Are Protective in Invasive Staphylococcus aureus Infection.
Authors: Brown A, Murphy A, Lalor S, Leech J, O'Keeffe K, Mac Aogain M, O'Halloran D, Lacey K, Tavakol M, Hearnden C, Fitzgerald-Hughes D, Humphreys H, Fennell J, van Wamel W, Foster T, Geoghegan J, Lavelle E, Rogers T, McLoughlin R
PLoS Pathog, 2015-11-05;11(11):e1005226.
Species: Mouse
Sample Types: Cell Culture Supernates
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Cholera toxin adjuvant promotes a balanced Th1/Th2/Th17 response independently of IL-12 and IL-17 by acting on Gsalpha in CD11b(+) DCs.
Authors: Mattsson J, Schon K, Ekman L, Fahlen-Yrlid L, Yrlid U, Lycke N
Mucosal Immunol, 2014-11-26;8(4):815-27.
Species: Mouse
Sample Types: Cell Culture Supernates
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The aryl hydrocarbon receptor: differential contribution to T helper 17 and T cytotoxic 17 cell development.
Authors: Hayes M, Ovcinnikovs V, Smith A, Kimber I, Dearman R
PLoS ONE, 2014-09-09;9(9):e106955.
Species: Mouse
Sample Types: Cell Culture Supernates
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Lymphotoxin beta receptor signaling limits mucosal damage through driving IL-23 production by epithelial cells.
Authors: Macho-Fernandez E, Koroleva E, Spencer C, Tighe M, Torrado E, Cooper A, Fu Y, Tumanov A
Mucosal Immunol, 2014-09-03;8(2):403-13.
Species: Mouse
Sample Types: Tissue Homogenates
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Unique macrophages different from M1/M2 macrophages inhibit T cell mitogenesis while upregulating Th17 polarization.
Authors: Tatano Y, Shimizu T, Tomioka H
Sci Rep, 2014-02-20;4(0):4146.
Species: Mouse
Sample Types: Cell Culture Supernates
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Exposure to atmospheric particulate matter enhances Th17 polarization through the aryl hydrocarbon receptor.
Authors: van Voorhis, Michael, Knopp, Samantha, Julliard, Walker, Fechner, John H, Zhang, Xiaoji, Schauer, James J, Mezrich, Joshua D
PLoS ONE, 2013-12-11;8(12):e82545.
Species: Mouse
Sample Types: Cell Culture Supernates
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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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Interleukin-23-mediated inflammation in Pseudomonas aeruginosa pulmonary infection.
Authors: Dubin PJ, Martz A, Eisenstatt JR, Fox MD, Logar A, Kolls JK
Infect. Immun., 2011-10-24;80(1):398-409.
Species: Mouse
Sample Types: BALF
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High expression of IL-22 suppresses antigen-induced immune responses and eosinophilic airway inflammation via an IL-10-associated mechanism.
Authors: Nakagome K, Imamura M, Kawahata K
J. Immunol., 2011-10-12;187(10):5077-89.
Species: Mouse
Sample Types: Tissue Homogenates
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The encephalitogenicity of T(H)17 cells is dependent on IL-1- and IL-23-induced production of the cytokine GM-CSF.
Authors: El-Behi M, Ciric B, Dai H
Nat. Immunol., 2011-04-24;12(6):568-75.
Species: Mouse
Sample Types: Cell Culture Supernates
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IL-1RL2 and its ligands contribute to the cytokine network in psoriasis.
Authors: Blumberg H, Dinh H, Dean C, Trueblood ES, Bailey K, Shows D, Bhagavathula N, Aslam MN, Varani J, Towne JE, Sims JE
J. Immunol., 2010-09-10;185(7):4354-62.
Species: Mouse
Sample Types: Tissue Homogenates
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