Human IL-33 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-33. 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-33
IL-33 (Interleukin-33) is released from physically damaged or necrotic cells. It triggers Th2-biased immune cell activation at sites of inflammation as well as regulatory T cell and M2 macrophage expansion. It additionally induces angiogenesis, promotes tumor cell migration and invasion, limits cardiac myocyte hypertrophy, and limits the development of atherosclerotic plaques. IL-33 signals through a receptor complex composed of ST2 and IL-1 RAcP. IL-1 RAcP also associates with IL-1 RI, IL-1 RII, IL-1 R6, and SCF R/c-kit. Soluble isoforms of ST2 and IL-1 RAcP function as decoy receptors that regulate IL-33 bioactivity.
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-33 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.
31
Citations: Showing 1 - 10
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Adiponectin and Interleukin-33: Possible Early Markers of Metabolic Syndrome
Authors: J Nesic, B Ljujic, V Rosic, A Djukic, M Rosic, I Petrovic, N Zornic, IP Jovanovic, S Petrovic, S Djukic
Journal of Clinical Medicine, 2022-12-24;12(1):.
Species: Human
Sample Types: Plasma
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Soluble ST2 and All-Cause Mortality in Patients with Chronic Obstructive Pulmonary Disease-A 10-Year Cohort Study
Authors: MH Urban, S Stojkovic, S Demyanets, C Hengstenbe, A Valipour, J Wojta, OC Burghuber
Journal of Clinical Medicine, 2021-12-23;11(1):.
Species: Human
Sample Types: Serum
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COVID-19 Modulates Inflammatory and Renal Markers That May Predict Hospital Outcomes among African American Males
Authors: W Fonseca, N Asai, K Yagi, CA Malinczak, G Savickas, CC Johnson, S Murray, EM Zoratti, NW Lukacs, J Li, CF Schuler Iv
Viruses, 2021-12-02;13(12):.
Species: Human
Sample Types: Whole Blood
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Adiponectin and Stnfr2 peripheral levels are associated with cardiovascular risk in patients with schizophrenia
Authors: ICS Dias, SM de Campos-, ELM Vieira, APL Mota, PS Azevedo, VTDS Anício, FC Guimarães, LM Mantovani, BF Cruz, AL Teixeira, JV Salgado
Journal of psychiatric research, 2021-11-09;0(0):.
Species: Human
Sample Types: Serum
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Soluble suppression of tumorigenicity 2 as outcome predictor after cardiopulmonary resuscitation: an observational prospective study
Authors: R Rezar, V Paar, C Seelmaier, I Pretsch, P Schwaiger, K Kopp, R Kaufmann, TK Felder, E Prinz, G Gemes, R Pistulli, UC Hoppe, B Wernly, M Lichtenaue
Scientific Reports, 2021-11-05;11(1):21756.
Species: Human
Sample Types: Serum
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PGC-1&alpha regulates airway epithelial barrier dysfunction induced by house dust mite
Authors: T Saito, T Ichikawa, T Numakura, M Yamada, A Koarai, N Fujino, K Murakami, S Yamanaka, Y Sasaki, Y Kyogoku, K Itakura, H Sano, K Takita, R Tanaka, T Tamada, M Ichinose, H Sugiura
Respiratory Research, 2021-02-19;22(1):63.
Species: Human
Sample Types: Cell Culture Supernates
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Graft IL-33 regulates infiltrating macrophages to protect against chronic rejection
Authors: T Li, Z Zhang, JG Bartolacci, GK Dwyer, Q Liu, L Mathews, M Velayutham, A Roessing, YC Lee, H Dai, S Shiva, MH Oberbarnsc, JL Dziki, SJ Mullett, SG Wendell, JD Wilkinson, SA Webber, MA Wood-Trage, SC Watkins, AJ Demetris, GS Hussey, SF Badylak, HR Turnquist
J. Clin. Invest., 2020-10-01;0(0):.
Species: Human
Sample Types: Serum
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Gene expression network analyses during infection with virulent and avirulent Trypanosoma cruzi�strains unveil a role for fibroblasts in neutrophil recruitment and activation
Authors: AER Oliveira, MCA Pereira, AT Belew, LRP Ferreira, LMN Pereira, EGA Neves, MDCP Nunes, BA Burleigh, WO Dutra, NM El-Sayed, RT Gazzinelli, SMR Teixeira
PLoS Pathog., 2020-08-18;16(8):e1008781.
Species: Human
Sample Types: Cell Culture Supernates
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The pro-inflammatory marker soluble suppression of tumorigenicity-2 (ST2) is reduced especially in diabetic morbidly obese patients undergoing bariatric surgery
Authors: S Demyanets, C Kaun, A Kaider, W Speidl, M Prager, S Oravec, P Hohensinne, J Wojta, G Rega-Kaun
Cardiovasc Diabetol, 2020-02-26;19(1):26.
Species: Human
Sample Types: Serum
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Identification of TNFR2 and IL-33 as therapeutic targets in localized fibrosis
Authors: D Izadi, TB Layton, L Williams, F McCann, M Cabrita, AI Espirito S, W Xie, M Fritzsche, H Colin-York, M Feldmann, KS Midwood, J Nanchahal
Sci Adv, 2019-12-04;5(12):eaay0370.
Species: Human
Sample Types: Cell Culture Supernates
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Helicobacter pylori Induces IL-33 Production and Recruits ST-2 to Lipid Rafts to Exacerbate Inflammation
Authors: CJ Kuo, CY Chen, HR Lo, CL Feng, HY Wu, MZ Huang, TN Liao, YA Chen, CH Lai
Cells, 2019-10-21;8(10):.
Species: Human
Sample Types: Cell Fraction
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Interleukin 1 Receptor-Like 1 (IL1RL1) Promotes Airway Bacterial and Viral Infection and Inflammation
Authors: N Schaunaman, A Sanchez, KG Dimasuay, N Pavelka, M Numata, R Alam, RJ Martin, HW Chu
Infect. Immun., 2019-06-20;87(7):.
Species: Human
Sample Types: Cell Culture Supernates
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Substance P and IL-33 administered together stimulate a marked secretion of IL-1? from human mast cells, inhibited by methoxyluteolin
Authors: A Taracanova, I Tsilioni, P Conti, ER Norwitz, SE Leeman, TC Theoharide
Proc. Natl. Acad. Sci. U.S.A., 2018-09-19;0(0):.
Species: Human
Sample Types: Serum
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The Effects of Varying Degree of MWCNT Carboxylation on Bioactivity in Various In Vivo and In Vitro Exposure Models
Authors: RF Hamilton, Z Wu, S Mitra, A Holian
Int J Mol Sci, 2018-01-25;19(2):.
Species: Human
Sample Types: Cell Culture Supernates
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Double-stranded RNA released from damaged articular chondrocytes promotes cartilage degeneration via Toll-like receptor 3-interleukin-33 pathway
Authors: C Li, K Chen, H Kang, Y Yan, K Liu, C Guo, J Qi, K Yang, F Wang, L Guo, C He, L Deng
Cell Death Dis, 2017-11-02;8(11):e3165.
Species: Human
Sample Types: Tissue Homogenates
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Predictive value of low interleukin-33 in critically ill patients
Authors: KA Krychtiuk, S Stojkovic, M Lenz, M Brekalo, K Huber, J Wojta, G Heinz, S Demyanets, WS Speidl
Cytokine, 2017-09-30;0(0):.
Species: Human
Sample Types: Serum
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IL33 and IL1RL1 variants are associated with asthma and atopy in a Brazilian population
Authors: GA Queiroz, RS Costa, NM Alcantara-, G Nunes de O, ML Barreto, VL Carneiro, CA Figueiredo
Int. J. Immunogenet, 2017-04-01;44(2):51-61.
Species: Human
Sample Types: Cell Culture Supernates
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IL-33/ST2 immune responses to respiratory bacteria in pediatric asthma
Authors: I Hentschke, A Graser, VO Melichar, A Kiefer, T Zimmermann, B Kro beta, P Haag, P Xepapadaki, NG Papadopoul, C Bogdan, S Finotto
Sci Rep, 2017-03-06;7(0):43426.
Species: Human
Sample Types: Tissue Secretion
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A Proinflammatory Role of Type 2 Innate Lymphoid Cells in Murine Immune-Mediated Hepatitis
Authors: Gisa Tiegs
J. Immunol., 2016-11-21;0(0):.
Species: Human
Sample Types: Plasma
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Rhinoviral stimuli, epithelial factors and ATP signalling contribute to bronchial smooth muscle production of IL-33.
Authors: Calven J, Akbarshahi H, Menzel M, Ayata C, Idzko M, Bjermer L, Uller L
J Transl Med, 2015-08-29;13(0):281.
Species: Human
Sample Types: Cell Culture Supernates
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Epidermal Expression and Regulation of Interleukin-33 during Homeostasis and Inflammation: Strong Species Differences.
Authors: Sundnes O, Pietka W, Loos T, Sponheim J, Rankin A, Pflanz S, Bertelsen V, Sitek J, Hol J, Haraldsen G, Khnykin D
J Invest Dermatol, 2015-03-04;135(7):1771-80.
Species: Human
Sample Types: Cell Lysates
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IL-33 is negatively associated with the BMI and confers a protective lipid/metabolic profile in non-diabetic but not diabetic subjects.
Authors: Hasan, Amal, Al-Ghimlas, Fahad, Warsame, Samia, Al-Hubail, Asma, Ahmad, Rasheed, Bennakhi, Abdullah, Al-Arouj, Monira, Behbehani, Kazem, Dehbi, Mohammed, Dermime, Said
BMC Immunol, 2014-05-10;15(0):19.
Species: Human
Sample Types: Serum
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Serum soluble ST2 is associated with ER-positive breast cancer.
Authors: Lu, Da-Peng, Zhou, Xiang-Yu, Yao, Lu-Tian, Liu, Cai-Gang, Ma, Wei, Jin, Feng, Wu, Yun-Fei
BMC Cancer, 2014-03-18;14(0):198.
Species: Human
Sample Types: Serum
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High plasma levels of soluble ST2 but not its ligand IL-33 is associated with severe forms of pediatric dengue.
Authors: Guerrero C, Arrieta A, Ramirez N, Rodriguez L, Vega R, Bosch I, Rodriguez J, Narvaez C, Salgado D
Cytokine, 2013-01-26;61(3):766-71.
Species: Human
Sample Types: Plasma
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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: Human
Sample Types: Cell Culture Supernates
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The role of IL-33 and its receptor ST2 in human nasal epithelium with allergic rhinitis.
Authors: Kamekura R, Kojima T, Takano K, Go M, Sawada N, Himi T
Clin. Exp. Allergy, 2012-02-01;42(2):218-28.
Species: Human
Sample Types: Serum
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ST2 and IL-33 in pregnancy and pre-eclampsia.
Authors: Granne I, Southcombe JH, Snider JV
PLoS ONE, 2011-09-16;6(9):e24463.
Species: Human
Sample Types: Cell Culture Supernates
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Soluble IL-1RII and IL-18 are associated with incipient upper extremity soft tissue disorders.
Authors: Rechardt M, Shiri R, Matikainen S, Viikari-Juntura E, Karppinen J, Alenius H
Cytokine, 2011-03-02;54(2):149-53.
Species: Human
Sample Types: Serum
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High Levels of Soluble ST2 and Low Levels of IL-33 in Sera of Patients with HIV Infection.
Authors: Miyagaki T, Sugaya M
J. Invest. Dermatol., 2010-12-09;131(0):794.
Species: Human
Sample Types: Serum
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Primary sources and immunological prerequisites for sST2 secretion in humans.
Authors: Mildner M, Storka A, Lichtenauer M
Cardiovasc. Res., 2010-04-02;87(4):769-77.
Species: Human
Sample Types: Cell Culture Supernates
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Expression of interleukin 1-like cytokine interleukin 33 and its receptor complex (ST2L and IL1RAcP) in human pancreatic myofibroblasts.
Authors: Nishida A, Andoh A, Imaeda H, Inatomi O, Shiomi H, Fujiyama Y
Gut, 2009-12-08;59(4):531-41.
Species: Human
Sample Types: Cell Culture Supernates
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