Human IL-33 Quantikine ELISA Kit

Newer Version Available: D3300B

Discontinued Product

D3300 has been discontinued and is replaced by D3300B.

Human IL-33 ELISA Cell Culture Supernate/Cell Lysate Standard Curve
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Human IL-33 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Cell Lysates (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL)
Sensitivity
1.65 pg/mL
Assay Range
6.3 - 400 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human IL-33
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human IL-33 Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-33 in cell culture supernates, cell lysates, serum, and plasma. It contains E. coli-expressed recombinant human IL-33 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human IL-33. Results obtained using natural human IL-33 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-33.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of a known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of a known concentration were tested in separate assays to determine inter-assay precision. Assays were performed by at least three technicians using two lots of kit components

Cell Culture Supernates, Cell Lysates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 34.1 112 214 36.4 116 224
Standard Deviation 1.21 3.22 5.36 2.8 6.3 10.4
CV% 3.5 2.9 2.5 7.7 5.4 4.6

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 34.3 111 218 39 120 230
Standard Deviation 1.45 3.33 5 2.2 5.6 12.8
CV% 4.2 3 2.3 5.6 4.7 5.6

Recovery

The recovery of human IL-33 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 95 81-106
EDTA Plasma (n=4) 86 82-92
Heparin Plasma (n=4) 84 80-90
Serum (n=4) 96 88-103

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human IL-33 were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Human IL-33 ELISA Linearity

Data Examples

Human IL-33 ELISA Cell Culture Supernate/Cell Lysate Standard Curve

Human IL-33 ELISA Serum/Plasma Standard Curve

Product Datasheets

Preparation and Storage

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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 100 µL Standard, Control, or Sample
  6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human IL-33 Quantikine ELISA Kit

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.

12 Citations: Showing 1 - 10
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  1. Genetic variants in IL-33/ST2 pathway with the susceptibility to hepatocellular carcinoma in a Chinese population
    Authors: ZH Wei, YY Li, SQ Huang, ZQ Tan
    Cytokine, 2018;0(0):.
    Species: Human
    Sample Types: Plasma
  2. Serum IL-33 level is associated with auto-antibodies but not with clinical response to biologic agents in rheumatoid arthritis
    Authors: E Rivière, J Sellam, J Pascaud, P Ravaud, JE Gottenberg, X Mariette
    Arthritis Res. Ther., 2018;20(1):122.
    Species: Human
    Sample Types: Serum
  3. In drug-induced, immune-mediated hepatitis, interleukin-33 reduces hepatitis and improves survival independently and as a consequence of FoxP3+ T-cell activity
    Authors: M Cottagiri, M Nyandjo, M Stephens, J Mantilla, H Saito, IR Mackay, NR Rose, DB Njoku
    Cell. Mol. Immunol., 2018;0(0):.
    Species: Human
    Sample Types: Serum
  4. Serum sST2 levels predict severe exacerbation of asthma
    Authors: M Watanabe, K Nakamoto, T Inui, M Sada, K Honda, M Tamura, Y Ogawa, T Yokoyama, T Saraya, D Kurai, H Ishii, H Takizawa
    Respir. Res., 2018;19(1):169.
    Species: Human
    Sample Types: Serum
  5. Upregulation of interleukin-33 and thymic stromal lymphopoietin levels in the lungs of idiopathic pulmonary fibrosis
    Authors: JU Lee, HS Chang, HJ Lee, CA Jung, DJ Bae, HJ Song, JS Park, ST Uh, YH Kim, KH Seo, CS Park
    BMC Pulm Med, 2017;17(1):39.
    Species: Human
    Sample Types: BALF
  6. Th1/Th17-Related Cytokines and Chemokines and Their Implications in the Pathogenesis of Pemphigus Vulgaris
    Authors: RP Timoteo, MV da Silva, CB Miguel, DA Silva, JD Catarino, V Rodrigues, H Sales-Camp, CJ Freire Oli
    Mediators Inflamm., 2017;2017(0):7151285.
    Species: Human
    Sample Types: Serum
  7. Immune and Imaging Correlates of Mild Cognitive Impairment Conversion to Alzheimer's Disease
    Authors: F La Rosa, M Saresella, F Baglio, F Piancone, I Marventano, E Calabrese, R Nemni, E Ripamonti, M Cabinio, M Clerici
    Sci Rep, 2017;7(1):16760.
    Species: Human
    Sample Types: Cell Culture Supernate
  8. Regulatory T cell frequency, but not plasma IL-33 levels, represents potential immunological biomarker to predict clinical response to intravenous immunoglobulin therapy
    Authors: MS Maddur, E Stephen-Vi, M Das, P Prakhar, VK Sharma, V Singh, M Rabin, J Trinath, KN Balaji, F Bolgert, JM Vallat, L Magy, SV Kaveri, J Bayry
    J Neuroinflammation, 2017;14(1):58.
    Species: Human
    Sample Types: Plasma
  9. Carotid plaque and bone density and microarchitecture in psoriatic arthritis: the correlation with soluble ST2
    Sci Rep, 2016;6(0):32116.
    Species: Human
    Sample Types: Plasma
  10. IL-33 reflects dynamics of disease activity in patients with autoimmune hemolytic anemia by regulating autoantibody production.
    Authors: Bu X, Zhang T, Wang C, Ren T, Wen Z
    J Transl Med, 2015;13(0):381.
    Species: Human
    Sample Types: Serum
  11. 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;61(3):766-71.
    Species: Human
    Sample Types: Plasma
  12. The challenge of measuring IL-33 in serum using commercial ELISA: lessons from asthma.
    Authors: Ketelaar M, Nawijn M, Shaw D, Koppelman G, Sayers I
    Clin Exp Allergy, 0;46(6):884-7.
    Species: Human
    Sample Types: Serum

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