Mouse/Rat IL-33 Quantikine ELISA Kit

  (19 citations)     
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Product Details
Assay Procedure
Citations (19)
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  • 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 (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
  • Sensitivity
    14.3 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse and rat 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 Mouse/Rat IL-33 immunoassay is a 4.5 hour solid-phase ELISA designed to measure IL-33 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse mature IL-33 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate both recombinant mouse and rat IL-33. Results obtained using natural IL-33 showed dose response 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 IL-33.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 41 41 41
Mean 58.2 298 630 57.6 282 599
Standard Deviation 5.6 17.4 33.5 5.89 16.3 30
CV% 9.6 5.8 5.3 10.2 5.8 5

Recovery

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

Sample Type Average % Recovery Range %
Mouse Cell Culture Supernates (n=4) 104 91-116
Mouse EDTA Plasma (n=4) 102 96-113
Mouse Heparin Plasma (n=4) 99 83-111
Mouse Serum (n=4) 90 81-103
Rat Cell Culture Supernates (n=4) 104 94-116
Rat EDTA Plasma (n=4) 93 83-100
Rat Heparin Plasma (n=4) 93 87-99
Rat Serum (n=4) 93 86-101
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of IL-33 in each matrix were diluted with Calibrator Diluent and assayed.
 IL-33 [HRP]
 IL-33 [HRP]
Product Datasheets

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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
Related Research Areas
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. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
Citations:

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.

19 Citations: Showing 1 - 10
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Species
Sample Type
  1. ?? T cells producing interleukin-17A regulate adipose regulatory T cell homeostasis and thermogenesis
    Authors: AC Kohlgruber, ST Gal-Oz, NM LaMarche, M Shimazaki, D Duquette, HN Nguyen, AI Mina, T Paras, A Tavakkoli, U von Andria, AS Banks, T Shay, MB Brenner, L Lynch
    Nat. Immunol., 2018;19(5):464-474.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  2. Characterization of the acute inflammatory profile and resolution of airway inflammation after Igf1r-gene targeting in a murine model of HDM-induced asthma
    Authors: S Piñeiro-He, E Alfaro-Arn, JA Gregory, R Torrens, C Ruíz-Martí, M Adner, IP López, JG Pichel
    PLoS ONE, 2017;12(12):e0190159.
    Species: Mouse
    Sample Type: Serum
  3. Stromal cell cadherin-11 regulates adipose tissue inflammation and diabetes
    Authors: SK Chang, AC Kohlgruber, F Mizoguchi, X Michelet, BJ Wolf, K Wei, PY Lee, L Lynch, D Duquette, V Ceperuelo-, AS Banks, MB Brenner
    J. Clin. Invest., 2017;0(0):.
    Species: Mouse
    Sample Type: Serum
  4. The PDGF-BB-SOX7 axis-modulated IL-33 in pericytes and stromal cells promotes metastasis through tumour-associated macrophages
    Nat Commun, 2016;7(0):11385.
    Species: Mouse
    Sample Type: Cell Lysates
  5. Membrane translocation of IL-33 receptor in ventilator induced lung injury.
    Authors: Yang S, Lin J, Wu S, Huang K, Jung F, Ma M, Wang Hsu G, Jow G
    PLoS ONE, 2015;10(3):e0121391.
    Species: Rat
    Sample Type: BALF
  6. Fibrates ameliorate the course of bacterial sepsis by promoting neutrophil recruitment via CXCR2.
    Authors: Tancevski I, Nairz M, Duwensee K, Auer K, Schroll A, Heim C, Feistritzer C, Hoefer J, Gerner R, Moschen A, Heller I, Pallweber P, Li X, Theurl M, Demetz E, Wolf A, Wolf D, Eller P, Ritsch A, Weiss G
    EMBO Mol Med, 2014;6(6):810-20.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  7. Impaired bone formation and increased osteoclastogenesis in mice lacking chemokine (C-C motif) ligand 5 (Ccl5).
    Authors: Wintges K, Beil F, Albers J, Jeschke A, Schweizer M, Claass B, Tiegs G, Amling M, Schinke T
    J Bone Miner Res, 2013;28(10):2070-80.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  8. An interleukin-33 gene polymorphism is a modifier for eosinophilia in rats.
    Authors: Luo H, Higuchi K, Matsumoto K, Mori M
    Genes Immun, 2013;14(3):192-7.
  9. Sirt2 suppresses inflammatory responses in collagen-induced arthritis.
    Authors: Lin J, Sun B, Jiang C, Hong H, Zheng Y
    Biochem Biophys Res Commun, 2013;441(4):897-903.
    Species: Mouse
    Sample Type: Serum
  10. IL-33 citrine reporter mice reveal the temporal and spatial expression of IL-33 during allergic lung inflammation.
    Authors: Hardman C, Panova V, McKenzie A
    Eur J Immunol, 2013;43(2):488-98.
    Species: Mouse
    Sample Type: BALF
  11. A novel method for procuring a large quantity of mature murine eosinophils in vivo.
    Authors: Kim W, Kim J, Shin C, Kim H, Doh Y, Cho HR, Kwon B
    J. Immunol. Methods, 2011;363(1):90-4.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  12. IL-33-activated dendritic cells are critical for allergic airway inflammation.
    Authors: Besnard AG, Togbe D, Guillou N, Erard F, Quesniaux V, Ryffel B
    Eur. J. Immunol., 2011;41(6):1675-86.
    Species: Mouse
    Sample Type: BALF
  13. IL-22 Is Produced by Innate Lymphoid Cells and Limits Inflammation in Allergic Airway Disease.
    Authors: Taube C, Tertilt C, Gyulveszi G, Dehzad N, Kreymborg K, Schneeweiss K, Michel E, Reuter S, Renauld JC, Arnold-Schild D, Schild H, Buhl R, Becher B
    PLoS ONE, 2011;6(7):e21799.
    Species: Mouse
    Sample Type: BALF
  14. IL-33-responsive lineage- CD25+ CD44(hi) lymphoid cells mediate innate type 2 immunity and allergic inflammation in the lungs.
    Authors: Bartemes KR, Iijima K, Kobayashi T, Kephart GM, McKenzie AN, Kita H
    J. Immunol., 2011;188(3):1503-13.
    Species: Mouse
    Sample Type: BALF
  15. IL-33 and M2a alveolar macrophages promote lung defense against the atypical fungal pathogen Pneumocystis murina.
    Authors: Nelson MP, Christmann BS, Werner JL, Metz AE, Trevor JL, Lowell CA, Steele C
    J. Immunol., 2011;186(4):2372-81.
    Species: Mouse
    Sample Type: Tissue Homogenates
  16. CpG oligodeoxynucleotides induce strong up-regulation of interleukin 33 via Toll-like receptor 9.
    Authors: Shimosato T, Fujimoto M, Tohno M, Sato T, Tateo M, Otani H, Kitazawa H
    Biochem. Biophys. Res. Commun., 2010;394(1):81-6.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  17. IL-33 is produced by mast cells and regulates IgE-dependent inflammation.
    Authors: Hsu CL, Neilsen CV, Bryce PJ
    PLoS ONE, 2010;5(8):e11944.
    Species: Mouse
    Sample Type: Tissue Homogenates
  18. IL-33 shifts macrophage polarization, promoting resistance against Pseudomonas aeruginosa keratitis.
    Authors: Hazlett LD, McClellan SA, Barrett RP, Huang X, Zhang Y, Wu M, Van Rooijen N, Szliter E
    Invest. Ophthalmol. Vis. Sci., 2010;51(3):1524-32.
    Species: Mouse
    Sample Type: Tissue Homogenates
  19. Identification of a serum-induced transcriptional signature associated with type 1 diabetes in the BioBreeding rat.
    Authors: Kaldunski M, Jia S, Geoffrey R, Basken J, Prosser S, Kansra S, Mordes JP, Lernmark A, Wang X, Hessner MJ
    Diabetes, 2010;59(10):2375-85.
    Species: Rat
    Sample Type: Serum
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