Human CCL11/Eotaxin Quantikine ELISA Kit DTX00: R&D Systems

Human CCL11/Eotaxin Quantikine ELISA Kit

(27 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    3.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Citrate Plasma (50 uL)
  • Sensitivity
    5 pg/mL
  • Assay Range
    15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Citrate Plasma)
  • Specificity
    Natural and recombinant human Eotaxin
  • 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.
Control Available
QC21, Quantikine Immunoassay Control Group 4 - Please Inquire
Product Summary
The Quantikine Human Eotaxin Immunoassay is a 3.5 hour solid phase ELISA designed to measure human Eotaxin in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human Eotaxin and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant human Eotaxin accurately. Results obtained measuring natural human eotaxin showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human Eotaxin.

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.
Serum, EDTA Plasma, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean64.510937763.4190400
Standard Deviation2.55.812.87.319.633.5
CV%3.95.33.411.510.38.4

Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean66.818439067.5193396
Standard Deviation28.423.57.818.240.2
CV%34.6611.69.410.2

Recovery

The recovery of Eotaxin spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 100 86-111
Citrate Plasma (n=5) 99 81-117
EDTA Plasma (n=5) 99 82-115
Serum (n=5) 95 83-109
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of Eotaxin in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human CCL11/Eotaxin Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CCL11/Eotaxin
CCL11/Eotaxin is a chemokine that interacts with CCR3 to induce eosinophil, basophil, dendritic cell, and Th2 cell chemoattraction during inflammatory and allergic reactions. Its production is upregulated in airway epithelium, alveolar macrophages, eosinophils, smooth muscle cells, chondrocytes, and fibroblasts following allergen or inflammatory challenge. CCL11 is also involved in the growth of myeloid cell progenitors and the differentiation of mast cells.
    • Entrez Gene IDs
      6356 (Human); 20292 (Mouse);
    • Alternate Names
      C-C motif chemokine 11; CCL11; chemokine (C-C motif) ligand 11; Eosinophil chemotactic protein; Eotaxin; eotaxin-1; MGC22554; SCYA11Small-inducible cytokine A11; small inducible cytokine subfamily A (Cys-Cys), member 11 (eotaxin);
    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. 100 µL Assay Diluent
    4.   Add 100 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 twice for a total of 3 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 1 hour.
    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. 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:

    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.

    27 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Profiling of cytokines, chemokines and other soluble proteins as a potential biomarker in colorectal cancer and polyps
      Authors: NA Johdi, L Mazlan, I Sagap, R Jamal
      Cytokine, 2017;99(0):35-42.
      Species: Human
      Sample Type: Serum
    2. Hydroxycarbamide reduces eosinophil adhesion and degranulation in sickle cell anaemia patients.
      Authors: Pallis F, Conran N, Fertrin K, Olalla Saad S, Costa F, Franco-Penteado C
      Br J Haematol, 2014;164(2):286-95.
      Species: Human
      Sample Type: Plasma
    3. Increased Eotaxin and MCP-1 Levels in Serum from Individuals with Periodontitis and in Human Gingival Fibroblasts Exposed to Pro-Inflammatory Cytokines.
      Authors: Bostrom E, Kindstedt E, Sulniute R, Palmqvist P, Majster M, Holm C, Zwicker S, Clark R, Onell S, Johansson I, Lerner U, Lundberg P
      PLoS ONE, 2013;10(8):e0134608.
      Species: Human
      Sample Type: Cell Culture Supernates
    4. Anti-inflammatory effects of methoxyphenolic compounds on human airway cells.
      Authors: Houser KR, Johnson DK, Ishmael FT
      J Inflamm (Lond), 2012;9(0):6.
      Species: Human
      Sample Type: Cell Culture Supernates
    5. Leukotriene D4 and interleukin-13 cooperate to increase the release of eotaxin-3 by airway epithelial cells.
      Authors: Provost V, Langlois A, Chouinard F, Rola-Pleszczynski M, Chakir J, Flamand N, Laviolette M
      PLoS ONE, 2012;7(8):e43544.
      Species: Human
      Sample Type: Cell Culture Supernates
    6. Opposing roles of IL-17A and IL-25 in the regulation of TSLP production in human nasal epithelial cells.
      Authors: Xu G, Zhang L, Wang DY, Xu R, Liu Z, Han DM, Wang XD, Zuo KJ, Li HB
      Allergy, 2010;65(5):581-9.
      Species: Human
      Sample Type: Cell Culture Supernates
    7. Presence of IL-5 protein and IgE antibodies to staphylococcal enterotoxins in nasal polyps is associated with comorbid asthma.
      Authors: Bachert C, Zhang N, Holtappels G, De Lobel L, Van Cauwenberge P, Liu S, Lin P, Bousquet J, Van Steen K
      J. Allergy Clin. Immunol., 2010;126(5):962.
      Species: Human
      Sample Type: Tissue Homogenates
    8. Eotaxins and CCR3 interaction regulates the Th2 environment of cutaneous T-cell lymphoma.
      Authors: Miyagaki T, Sugaya M, Fujita H
      J. Invest. Dermatol., 2010;130(9):2304-11.
      Species: Human
      Sample Type: Serum
    9. Staphylococcus aureus in nasal lavage and biopsy of patients with chronic rhinosinusitis.
      Authors: Niederfuhr A, Kirsche H, Deutschle T, Poppert S, Riechelmann H, Wellinghausen N
      Allergy, 2008;63(10):1359-67.
      Species: Human
      Sample Type: Nasal Lavage
    10. Infiltrated neutrophils acquire novel chemokine receptor expression and chemokine responsiveness in chronic inflammatory lung diseases.
      Authors: Hartl D, Krauss-Etschmann S, Koller B, Hordijk PL, Kuijpers TW, Hoffmann F, Hector A, Eber E, Marcos V, Bittmann I, Eickelberg O, Griese M, Roos D
      J. Immunol., 2008;181(11):8053-67.
      Species: Human
      Sample Type: BALF
    11. Circulating thymus- and activation-regulated chemokine/CCL17 is a useful biomarker for discriminating acute eosinophilic pneumonia from other causes of acute lung injury.
      Authors: Miyazaki E, Nureki S, Ono E, Ando M, Matsuno O, Fukami T, Ueno T, Kumamoto T
      Chest, 2007;131(6):1726-34.
      Species: Human
      Sample Type: BALF
    12. Cerebrospinal fluid eotaxin and eotaxin-2 levels in human eosinophilic meningitis associated with angiostrongyliasis.
      Authors: Intapan PM, Niwattayakul K, Sawanyawisuth K, Chotmongkol V, Maleewong W
      Cytokine, 2007;39(2):138-41.
      Species: Human
      Sample Type: CSF
    13. Computed tomographic scan-diagnosed chronic obstructive pulmonary disease-emphysema: eotaxin-1 is associated with bronchodilator response and extent of emphysema.
      Authors: Miller M, Ramsdell J, Friedman PJ, Cho JY, Renvall M, Broide DH
      J. Allergy Clin. Immunol., 2007;120(5):1118-25.
      Species: Human
      Sample Type: BALF
    14. Effect of IL-13 receptor alpha2 levels on the biological activity of IL-13 variant R110Q.
      Authors: Andrews AL, Bucchieri F, Arima K, Izuhara K, Holgate ST, Davies DE, Holloway JW
      J. Allergy Clin. Immunol., 2007;120(1):91-7.
      Species: Human
      Sample Type: Cell Culture Supernates
    15. Effect of IL-6 trans-signaling on the pro-remodeling phenotype of airway smooth muscle.
      Authors: Ammit AJ, Moir LM, Oliver BG, Hughes JM, Alkhouri H, Ge Q, Burgess JK, Black JL, Roth M
      Am. J. Physiol. Lung Cell Mol. Physiol., 2007;292(1):L199-206.
      Species: Human
      Sample Type: Cell Culture Supernates
    16. Systemic responses after bronchial aspirin challenge in sensitive patients with asthma.
      Authors: Makowska JS, Grzegorczyk J, Bienkiewicz B, Wozniak M, Kowalski ML
      J. Allergy Clin. Immunol., 2007;121(2):348-54.
      Species: Human
      Sample Type: Serum
    17. Relationship between eosinophilia and levels of chemokines (CCL5 and CCL11) and IL-5 in bronchoalveolar lavage fluid of patients with mustard gas-induced pulmonary fibrosis.
      Authors: Emad A, Emad Y
      J. Clin. Immunol., 2007;27(6):605-12.
      Species: Human
      Sample Type: BALF
    18. Mast cell beta-tryptase selectively cleaves eotaxin and RANTES and abrogates their eosinophil chemotactic activities.
      Authors: Pang L, Nie M, Corbett L, Sutcliffe A, Knox AJ
      J. Immunol., 2006;176(6):3788-95.
      Species: Human
      Sample Type: Cell Culture Supernates
    19. Markers of eosinophilic inflammation and risk prediction in patients with coronary artery disease.
      Authors: Falcone C, Minoretti P, D&amp;amp;apos;Angelo A, Buzzi MP, Coen E, Emanuele E, Aldeghi A, Olivieri V, Geroldi D
      Eur. J. Clin. Invest., 2006;36(4):211-7.
      Species: Human
      Sample Type: Plasma
    20. Regulatory mechanisms of galectin-9 and eotaxin-3 synthesis in epidermal keratinocytes: possible involvement of galectin-9 in dermal eosinophilia of Th1-polarized skin inflammation.
      Authors: Igawa K, Satoh T, Hirashima M, Yokozeki H
      Allergy, 2006;61(12):1385-91.
      Species: Human
      Sample Type: Cell Culture Supernates
    21. IL-13 receptor alpha 2: a regulator of IL-13 and IL-4 signal transduction in primary human fibroblasts.
      Authors: Andrews AL, Nasir T, Bucchieri F, Holloway JW, Holgate ST, Davies DE
      J. Allergy Clin. Immunol., 2006;118(4):858-65.
      Species: Human
      Sample Type: Cell Culture Supernates
    22. Airway inflammation during stable and acutely exacerbated chronic obstructive pulmonary disease.
      Authors: Fujimoto K, Yasuo M, Urushibata K, Hanaoka M, Koizumi T, Kubo K
      Eur. Respir. J., 2005;25(4):640-6.
      Species: Human
      Sample Type: Sputum
    23. Respiratory syncytial virus and other respiratory viruses during the first 3 months of life promote a local TH2-like response.
      Authors: Kristjansson S, Bjarnarson SP, Wennergren G, Palsdottir AH, Arnadottir T, Haraldsson A, Jonsdottir I
      J. Allergy Clin. Immunol., 2005;116(4):805-11.
      Species: Human
      Sample Type: Tissue Secretion
    24. Enhanced monocyte chemoattractant protein-3/CC chemokine ligand-7 in usual interstitial pneumonia.
      Authors: Choi ES, Jakubzick C, Carpenter KJ, Kunkel SL, Evanoff H, Martinez FJ, Flaherty KR, Toews GB, Colby TV, Kazerooni EA, Gross BH, Travis WD, Hogaboam CM
      Am. J. Respir. Crit. Care Med., 2004;170(5):508-15.
      Species: Human
      Sample Type: Cell Culture Supernates
    25. Regulation of the interleukin-5 receptor alpha-subunit on peripheral blood eosinophils from healthy subjects.
      Authors: Hellman C, Hallden G, Hylander B, Lundahl J
      Clin. Exp. Immunol., 2003;131(1):75-81.
      Species: Human
      Sample Type: Peritoneal Fluid
    26. CCR5-binding chemokines modulate CXCL12 (SDF-1)-induced responses of progenitor B cells in human bone marrow through heterologous desensitization of the CXCR4 chemokine receptor.
      Authors: Honczarenko M, Le Y, Glodek AM, Silberstein LE
      Blood, 2002;100(7):2321-9.
      Species: Human
      Sample Type: Cell Culture Supernates
    27. CCL11 is increased in the CNS in chronic traumatic encephalopathy but not in Alzheimer's disease.
      Authors: Cherry J, Stein T, Tripodis Y, Alvarez V, Huber B, Au R, Kiernan P, Daneshvar D, Mez J, Solomon T, Alosco M, McKee A
      PLoS ONE, 0;12(9):e0185541.
      Species: Human
      Sample Type: CSF
    Expand to show all 27 Citations
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