Human IL-33 DuoSet ELISA

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Ancillary Products Available
Human Fucose Detection ELISA Standard Curve
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Product Details
Citations (18)
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Human IL-33 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sufficient Materials
For fifteen 96-well plates*
Please see the product datasheet

* 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 Interleukin 33 (IL-33). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma 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

Block Buffer: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Reagent Diluent: 0.1% BSA, 0.05% Tween 20 in Tris-buffered Saline (20 mM Trizma base, 150 mM NaCI) 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)

Data Example

Human Fucose Detection ELISA Standard Curve

Product Datasheets

Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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.

Long Name:
Interleukin 33
Entrez Gene IDs:
90865 (Human); 77125 (Mouse)
Alternate Names:
C9orf26; C9orf26chromosome 9 open reading frame 26 (NF-HEV); DKFZp586H0523; DVS27; DVS27-related protein; IL1F11; IL-1F11; IL33; IL-33; interleukin 33; Interleukin-1 family member 11; interleukin-33; NFHEV; NF-HEV; NF-HEVNFEHEV; Nuclear factor from high endothelial venules; RP11-575C20.2

Assay Procedure


Plate Preparation

  1. 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.
  2. 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.
  3. Block plates by adding 300 μL of Block Buffer to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. 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.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. 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.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. 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.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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.

18 Citations: Showing 1 - 10
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  1. 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;0(0):.
    Species: Human
    Sample Types: Serum
  2. 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;19(2):.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. 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;8(11):e3165.
    Species: Human
    Sample Types: Tissue Homogenates
  4. 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;7(0):43426.
    Species: Human
    Sample Types: Tissue Secretion
  5. 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;44(2):51-61.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. 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;0(0):.
    Species: Human
    Sample Types: Serum
  7. A Proinflammatory Role of Type 2 Innate Lymphoid Cells in Murine Immune-Mediated Hepatitis
    Authors: Gisa Tiegs
    J. Immunol., 2016;0(0):.
    Species: Human
    Sample Types: Plasma
  8. 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;135(7):1771-80.
    Species: Human
    Sample Types: Cell Lysates
  9. 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;13(0):281.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. 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;14(0):198.
    Species: Human
    Sample Types: Serum
  11. 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;15(0):19.
    Species: Human
    Sample Types: Serum
  12. Interleukin-1alpha controls allergic sensitization to inhaled house dust mite via the epithelial release of GM-CSF and IL-33.
    J. Exp. Med., 2012;209(8):1505-17.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. 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;42(2):218-28.
    Species: Human
    Sample Types: Serum
  14. ST2 and IL-33 in pregnancy and pre-eclampsia.
    Authors: Granne I, Southcombe JH, Snider JV
    PLoS ONE, 2011;6(9):e24463.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. 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;54(2):149-53.
    Species: Human
    Sample Types: Serum
  16. 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, 2010;59(4):531-41.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Primary sources and immunological prerequisites for sST2 secretion in humans.
    Authors: Mildner M, Storka A, Lichtenauer M
    Cardiovasc. Res., 2010;87(4):769-77.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. 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;131(0):794.
    Species: Human
    Sample Types: Serum


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Human IL-33 DuoSet ELISA
By Anonymous on 09/16/2019
Sample Tested: Cell culture supernatant

Human IL-33 DuoSet ELISA
By Anonymous on 08/22/2019
Sample Tested: EDTA Plasma

Human IL-33 DuoSet ELISA
By Anonymous on 08/11/2017
Sample Tested: human serum