Human 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 human 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 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-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 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-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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Interferon subverts an AHR-JUN axis to promote CXCL13+ T cells in lupus
Authors: Law, C;Wacleche, VS;Cao, Y;Pillai, A;Sowerby, J;Hancock, B;Horisberger, A;Bracero, S;Skidanova, V;Li, Z;Adejoorin, I;Dillon, E;Benque, IJ;Nunez, DP;Simmons, DP;Keegan, J;Chen, L;Baker, T;Brohawn, PZ;Al-Mossawi, H;Hao, LY;Jones, B;Rao, N;Qu, Y;Alves, SE;Accelerating Medicines Partnership: RA/SLE Network, ;Jonsson, AH;Shaw, KS;Vleugels, RA;Massarotti, E;Costenbader, KH;Brenner, MB;Lederer, JA;Hultquist, JF;Choi, J;Rao, DA;
Nature
Species: Human
Sample Types: Cell Culture Supernates
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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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Mild Cognitive Impairment Is Associated with Enhanced Activation of Th17 Lymphocytes in Non-Alcoholic Fatty Liver Disease
Authors: Fiorillo, A;Gallego, JJ;Casanova-Ferrer, F;Giménez-Garzó, C;Urios, A;Ballester, MP;Durbán, L;Rios, MP;Megías, J;San Miguel, T;Kosenko, E;Escudero-García, D;Benlloch, S;Felipo, V;Montoliu, C;
International journal of molecular sciences
Species: Human
Sample Types: Plasma
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Mild Cognitive Impairment Is Associated with Enhanced Activation of Th17 Lymphocytes in Non-Alcoholic Fatty Liver Disease
Authors: Fiorillo, A;Gallego, JJ;Casanova-Ferrer, F;Giménez-Garzó, C;Urios, A;Ballester, MP;Durbán, L;Rios, MP;Megías, J;San Miguel, T;Kosenko, E;Escudero-García, D;Benlloch, S;Felipo, V;Montoliu, C;
International journal of molecular sciences
Species: Human
Sample Types: Cell Culture Supernates, Plasma
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Effect of exogenous IL-37 on immune cells from a patient carrying a potential IL37 loss-of-function variant: A case study
Authors: LU Teufel, CI van der Ma, V Klück, A Simons, A Hoischen, V Vernimmen, LAB Joosten, RJW Arts
Cytokine, 2022-12-05;162(0):156102.
Species: Human
Sample Types: Cell Culture Supernates
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IL-17A in Human Liver: Significant Source of Inflammation and Trigger of Liver Fibrosis Initiation.
Authors: Kartasheva-Ebertz D, Gaston J, Lair-Mehiri L, Mottez E, Buivan T, Massault P, Scatton O, Gaujoux S, Vaillant J, Pol S, Lagaye S
Int J Mol Sci, 2022-08-29;23(17):.
Species: Human
Sample Types: Tissue Culture Supernates
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A novel Streptococcus pneumoniae human challenge model demonstrates Treg lymphocyte recruitment to the infection site
Authors: G Szylar, R Wysoczansk, H Marshall, DJB Marks, R José, ME Ehrenstein, JS Brown
Scientific Reports, 2022-03-07;12(1):3990.
Species: Human
Sample Types: Blister Fluid
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Reduced Interleukin-17-Expressing Cells in Cutaneous Melanoma
Authors: A Tosi, L Nardinocch, ML Carbone, L Capriotti, E Pagani, S Mastroeni, C Fortes, F Scopelliti, C Cattani, F Passarelli, A Rosato, S D'Atri, CM Failla, A Cavani
Biomedicines, 2021-12-16;9(12):.
Species: Human
Sample Types: Cell Culture Supernates
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Tofacitinib-Induced Modulation of Intestinal Adaptive and Innate Immunity and Factors Driving Cellular and Systemic Pharmacokinetics
Authors: B Texler, A Zollner, V Reinstadle, SJ Reider, S Macheiner, B Jelusic, A Pfister, C Watschinge, N Przysiecki, H Tilg, H Oberacher, AR Moschen
Cellular and Molecular Gastroenterology and Hepatology, 2021-10-06;0(0):.
Species: Human
Sample Types: Cell culture supernate
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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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Diet influences the functions of the human intestinal microbiome
Authors: M De Angelis, I Ferrocino, FM Calabrese, F De Filippi, N Cavallo, S Siragusa, S Rampelli, R Di Cagno, K Rantsiou, L Vannini, N Pellegrini, C Lazzi, S Turroni, N Lorusso, M Ventura, M Chieppa, E Neviani, P Brigidi, PW O'Toole, D Ercolini, M Gobbetti, L Cocolin
Sci Rep, 2020-03-06;10(1):4247.
Species: Human
Sample Types: Cell Culture Supernates
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Ex vivo culture of lesional psoriasis skin for pharmacological testing
Authors: ML Tiirikaine, A Woetmann, H Norsgaard, LF Santamaria, P Lovato
J. Dermatol. Sci., 2019-12-27;0(0):.
Species: Human
Sample Types: Tissue Lysates
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PASylation of IL-1 Receptor antagonist (IL-1Ra) retains IL-1 blockade and extends its duration in mouse urate crystal-induced peritonitis
Authors: NE Powers, B Swartzwelt, C Marchetti, DM de Graaf, A Lerchner, M Schlapschy, R Datar, U Binder, CK Edwards, A Skerra, CA Dinarello
J. Biol. Chem., 2019-12-09;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Lysosomotropic beta blockers induce oxidative stress and IL23A production in Langerhans cells
Authors: G Müller, C Lübow, G Weindl
Autophagy, 2019-11-07;0(0):1-16.
Species: Human
Sample Types: Cell Culture Supernates
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In Vitro Induction of T Helper 17 Cells by Synergistic Activation of Human Monocyte-Derived Langerhans Cell-Like Cells with Bacterial Agonists
Authors: R Gramlich, E Aliahmadi, M Peiser
Int J Mol Sci, 2019-03-19;20(6):.
Species: Human
Sample Types: Cell Culture Supernates
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Inactivation of the interleukin-22 pathway in the airways of cystic fibrosis patients
Authors: A Guillon, D Brea, E Luczka, V Hervé, S Hasanat, C Thorey, M Pérez-Cruz, J Hordeaux, J Mankikian, P Gosset, C Coraux, M Si-Tahar
Cytokine, 2018-10-28;0(0):.
Species: Human
Sample Types: Sputum
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Cross-Tissue Transcriptomic Analysis of Human Secondary Lymphoid Organ-Residing ILC3s Reveals a Quiescent State in the Absence of Inflammation
Authors: YE Bar-Ephrai, F Cornelisse, N Papazian, T Konijn, RM Hoogenboez, MA Sanders, BA Westerman, M Gönültas, J Kwekkeboom, JMM Den Haan, RM Reijmers, RE Mebius, T Cupedo
Cell Rep, 2017-10-17;21(3):823-833.
Species: Human
Sample Types: Cell Culture Supernates
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TB lymphadenitis is associated with enhanced baseline and antigen - specific induction of Type 1 and Type 17 cytokines and reduced IL-1? and IL-18 at the site of infection
Authors: GR Kathamuthu, K Moideen, D Baskaran, VV Banurekha, D Nair, G Sekar, R Sridhar, B Vidyajayan, G Gajendrara, DK Parandhama, A Srinivasan, S Babu
Clin. Vaccine Immunol, 2017-05-05;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Activation of Immune and Defense Responses in the Intestinal Mucosa by Outer Membrane Vesicles of Commensal and Probiotic Escherichia coli Strains
Authors: MJ Fábrega, L Aguilera, R Giménez, E Varela, M Alexandra, M Antolín, J Badía, L Baldomà
Front Microbiol, 2016-05-11;7(0):705.
Species: Human
Sample Types: Cell Culture Supernates
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Bilberry-Derived Anthocyanins Modulate Cytokine Expression in the Intestine of Patients with Ulcerative Colitis
PLoS ONE, 2016-05-06;11(5):e0154817.
Species: Human
Sample Types: Serum
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Ultraviolet light converts propranolol, a nonselective beta-blocker and potential lupus-inducing drug, into a proinflammatory AhR ligand.
Authors: Dorgham K, Amoura Z, Parizot C, Arnaud L, Frances C, Pionneau C, Devilliers H, Pinto S, Zoorob R, Miyara M, Larsen M, Yssel H, Gorochov G, Mathian A
Eur J Immunol, 2015-10-16;45(11):3174-87.
Species: Human
Sample Types: Cell Culture Supernates
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Association of Mucosal Organisms with Patterns of Inflammation in Chronic Rhinosinusitis.
Authors: Chalermwatanachai T, Zhang N, Holtappels G, Bachert C
PLoS ONE, 2015-08-14;10(8):e0136068.
Species: Human
Sample Types: Tissue Homogenates
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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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Increased expression of IL-22 is associated with disease activity in Behcet's disease.
Authors: Cai, Tao, Wang, Qian, Zhou, Qingyun, Wang, Chaokui, Hou, Shengpin, Qi, Jian, Kijlstra, Aize, Yang, Peizeng
PLoS ONE, 2013-03-19;8(3):e59009.
Species: Human
Sample Types: Cell Culture Supernates
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Susceptibility-associated genetic variation at IL12B enhances Th1 polarization in psoriasis.
Authors: Johnston A, Xing X, Swindell W, Kochkodan J, Riblett M, Nair R, Stuart P, Ding J, Voorhees J, Elder J, Gudjonsson J
Hum Mol Genet, 2013-01-31;22(9):1807-15.
Species: Human
Sample Types: Serum
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IL-38 binds to the IL-36 receptor and has biological effects on immune cells similar to IL-36 receptor antagonist.
Authors: van de Veerdonk FL, Stoeckman AK, Wu G, Boeckermann AN, Azam T, Netea MG, Joosten LA, Van der Meer JW, Hao R, Kalabokis V, Dinarello CA
Proc. Natl. Acad. Sci. U.S.A., 2012-02-06;109(8):3001-5.
Species: Human
Sample Types: Cell Culture Supernates
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Interleukin-21 induces the differentiation of human Tc22 cells via phosphorylation of signal transducers and activators of transcription.
Authors: Liu Y, Yang B, Ma J, Wang H, Huang F, Zhang J, Chen H, Wu C
Immunology, 2011-01-07;132(4):540-8.
Species: Human
Sample Types: Cell Culture Supernates
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IL-17 amplifies human contact hypersensitivity by licensing hapten nonspecific Th1 cells to kill autologous keratinocytes.
Authors: Pennino D, Eyerich K, Scarponi C, Carbone T, Eyerich S, Nasorri F, Garcovich S, Traidl-Hoffmann C, Albanesi C, Cavani A
J. Immunol., 2010-03-31;184(9):4880-8.
Species: Human
Sample Types: Cell Culture Supernates
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Chronic mucocutaneous candidiasis in APECED or thymoma patients correlates with autoimmunity to Th17-associated cytokines.
Authors: Kisand K, Boe Wolff AS, Podkrajsek KT, Tserel L, Link M, Kisand KV, Ersvaer E, Perheentupa J, Erichsen MM, Bratanic N, Meloni A, Cetani F, Perniola R, Ergun-Longmire B, Maclaren N, Krohn KJ, Pura M, Schalke B, Strobel P, Leite MI, Battelino T, Husebye ES, Peterson P, Willcox N, Meager A
J. Exp. Med., 2010-02-01;207(2):299-308.
Species: Human
Sample Types: Cell Culture Supernates
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Protective immunity to systemic infection with attenuated Salmonella enterica serovar enteritidis in the absence of IL-12 is associated with IL-23-dependent IL-22, but not IL-17.
Authors: Schulz SM, Kohler G, Schutze N, Knauer J, Straubinger RK, Chackerian AA, Witte E, Wolk K, Sabat R, Iwakura Y, Holscher C, Muller U, Kastelein RA, Alber G
J. Immunol., 2008-12-01;181(11):7891-901.
Species: Mouse
Sample Types: Serum
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We used this kit for the quantification of IL22 in human serum and plasma. We could not see any signal in our samples. Nevertheless, the standard curve works very well and well-described protocol.