Human IL-5 Quantikine ELISA Kit D5000B: R&D Systems
  • 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), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL), Urine (100 uL)
  • Sensitivity
    1.08 pg/mL
  • Assay Range
    3.9 - 250 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
  • Specificity
    Natural and recombinant human IL-5
  • 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
QC20, Quantikine Immunoassay Control Group 3 - Please Inquire
Product Summary
The Quantikine Human IL-5 Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-5 levels in cell culture supernates, serum, plasma, and urine. It contains Sf 21-expressed recombinant human IL-5 and antibodies raised against the recombinant factor and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IL-5 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 IL-5.

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, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean31.395.617332.997.9180
Standard Deviation1.43.56.546.78.4
CV%4.53.73.812.26.84.7

Cell Culture Supernates, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean28.189.317031.292.6174
Standard Deviation0.8927.272.54.28.7
CV%3.22.24.284.55

Recovery

The recovery of human IL-5 spiked to different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 101 95-107
EDTA Plasma (n=4) 104 86-118
Heparin Plasma (n=4) 101 87-112
Serum (n=4) 104 88-116
Urine (n=4) 96 91-106
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of IL-5 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human IL-5 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-5
IL-5 (Interleukin-5) is a secreted cytokine that is expressed as a covalent dimer. It is produced by activated Th2 cells, eosinophils, mast cells, and iNKT cells and promotes eosinophilic and basophilic differentiation and activation. IL-5 signals through a receptor complex containing IL-5 R alpha and the Common beta Chain/CD131 on eosinophils and basophils. Naturally occurring soluble forms of IL-5 R alpha retain the ability to bind IL-5 and limit its bioavailability.
    • Long Name
      Interleukin 5
    • Entrez Gene IDs
      3567 (Human); 16191 (Mouse); 24497 (Rat); 397409 (Porcine); 280825 (Bovine); 403790 (Canine); 493803 (Feline);
    • Alternate Names
      BCDF mu; B-cell differentiation factor I; BCGFII; EDF; Eo-CSF; Eosinophil differentiation factor; IL5; IL-5; IL-5T-cell replacing factor; interleukin 5 (colony-stimulating factor, eosinophil); interleukin-5; TRF; TRFB cell differentiation factor I;
    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. 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:

    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
    Filter your results:

    Species
    Sample Type
    1. Live attenuated B. pertussis BPZE1 rescues the immune functions of Respiratory Syncytial virus infected human dendritic cells by promoting Th1/Th17 responses.
      Authors: Schiavoni I, Fedele G, Quattrini A, Bianco M, Schnoeller C, Openshaw P, Locht C, Ausiello C
      PLoS ONE, 2014;9(6):e100166.
      Species: Human
      Sample Type: Cell Culture Supernates
    2. Immunoepidemiological profiling of onchocerciasis patients reveals associations with microfilaria loads and ivermectin intake on both individual and community levels.
      Authors: Arndts, Kathrin, Specht, Sabine, Debrah, Alexande, Tamarozzi, Francesc, Klarmann Schulz, Ute, Mand, Sabine, Batsa, Linda, Kwarteng, Alexande, Taylor, Mark, Adjei, Ohene, Martin, Coralie, Layland, Laura E, Hoerauf, Achim
      PLoS Negl Trop Dis, 2014;8(2):e2679.
      Species: Human
      Sample Type: Plasma
    3. Anti-interleukin-5 and multiple autoantibodies are associated with human atherosclerotic diseases and serum interleukin-5 levels.
      Authors: Ishigami, Tomoaki, Abe, Kaito, Aoki, Ichiro, Minegishi, Shintaro, Ryo, Akihide, Matsunaga, Satoko, Matsuoka, Kazuhiro, Takeda, Hiroyuki, Sawasaki, Tatsuya, Umemura, Satoshi, Endo, Yaeta
      FASEB J, 2013;27(9):3437-45.
      Species: Human
      Sample Type: Serum
    4. 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
    5. Effects of atorvastatin added to inhaled corticosteroids on lung function and sputum cell counts in atopic asthma.
      Authors: Hothersall EJ, Chaudhuri R, McSharry C, Donnelly I, Lafferty J, McMahon AD, Weir CJ, Meiklejohn J, Sattar N, McInnes I, Wood S, Thomson NC
      Thorax, 2008;63(12):1070-5.
      Species: Human
      Sample Type: Serum
    6. 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
    7. Detection of pancreatic cancer using antibody microarray-based serum protein profiling.
      Authors: Ingvarsson J, Wingren C, Carlsson A, Ellmark P, Wahren B, Engstrom G, Harmenberg U, Krogh M, Peterson C, Borrebaeck CA
      Proteomics, 2008;8(11):2211-9.
      Species: Human
      Sample Type: Serum
    8. Mycobacterium bovis Bacillus Calmette-Guerin suppresses inflammatory Th2 responses by inducing functional alteration of TSLP-activated dendritic cells.
      Authors: Yokoi T, Amakawa R, Tanijiri T, Sugimoto H, Torii Y, Amuro H, Son Y, Tajima K, Liu YJ, Ito T, Fukuhara S
      Int. Immunol., 2008;20(10):1321-9.
      Species: Human
      Sample Type: Cell Culture Supernates
    9. Distinct subsets of human invariant NKT cells differentially regulate T helper responses via dendritic cells.
      Authors: Liu TY, Uemura Y, Suzuki M, Narita Y, Hirata S, Ohyama H, Ishihara O, Matsushita S
      Eur. J. Immunol., 2008;38(4):1012-23.
      Species: Human
      Sample Type: Cell Culture Supernates
    10. 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
    11. 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
    12. 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
    13. 1, 25 Dihydroxyvitamin D3 modulated cytokine response in pulmonary tuberculosis.
      Authors: Vidyarani M, Selvaraj P, Jawahar MS, Narayanan PR
      Cytokine, 2007;40(2):128-34.
      Species: Human
      Sample Type: Cell Culture Supernates
    14. Placental growth factor down-regulates type 1 T helper immune response by modulating the function of dendritic cells.
      Authors: Lin YL, Liang YC, Chiang BL
      J. Leukoc. Biol., 2007;82(6):1473-80.
      Species: Human
      Sample Type: Cell Culture Supernates
    15. T-lymphocyte activation and the levels of eosinophilic cationic protein and interleukin-5 in asthmatic children with acute exacerbation and effect of glucocorticoid treatment.
      Authors: Kocak AK, Bor O, Yildiz B, Erdogan L, Us T
      Allergy Asthma Proc, 2006;27(4):371-7.
      Species: Human
      Sample Type: Serum
    16. A network-based analysis of the late-phase reaction of the skin.
      Authors: Benson M, Langston MA, Adner M, Andersson B, Torinssson-Naluai A, Cardell LO
      J. Allergy Clin. Immunol., 2006;118(1):220-5.
      Species: Human
      Sample Type: Cell Culture Supernates
    17. Chemokines indicate allergic bronchopulmonary aspergillosis in patients with cystic fibrosis.
      Authors: Hartl D, Latzin P, Zissel G, Krane M, Krauss-Etschmann S, Griese M
      Am. J. Respir. Crit. Care Med., 2006;173(12):1370-6.
      Species: Human
      Sample Type: Serum
    18. Relationships between allergic inflammation and nasal airflow in children with seasonal allergic rhinitis.
      Authors: Ciprandi G, Tosca MA, Marseglia GL, Klersy C
      Ann. Allergy Asthma Immunol., 2005;94(2):258-61.
      Species: Human
      Sample Type: Nasal Lavage
    19. Eotaxin-3 and interleukin-5 pleural fluid levels are associated with pleural fluid eosinophilia in post-coronary artery bypass grafting pleural effusions.
      Authors: Kalomenidis I, Stathopoulos GT, Barnette R, Guo Y, Peebles RS, Blackwell TS, Light RW
      Chest, 2005;127(6):2094-100.
      Species: Human
      Sample Type: Pleural Effusion
    20. 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
    21. Effects of antiasthmatic agents on the functions of peripheral blood monocyte-derived dendritic cells from atopic patients.
      Authors: Saeki S, Matsuse H, Kondo Y, Machida I, Kawano T, Tomari S, Obase Y, Fukushima C, Kohno S
      J. Allergy Clin. Immunol., 2004;114(3):538-44.
      Species: Human
      Sample Type: Cell Culture Supernates
    22. Effect of ozone exposure on airway responses to inhaled allergen in asthmatic subjects.
      Authors: Chen LL, Tager IB, Peden DB, Christian DL, Ferrando RE, Welch BS, Balmes JR
      Chest, 2004;125(6):2328-35.
    23. Antifungal immune reactivity in nasal polyposis.
      Authors: Pitzurra L, Bellocchio S, Nocentini A, Bonifazi P, Scardazza R, Gallucci L, Stracci F, Simoncelli C, Bistoni F, Romani L
      Infect. Immun., 2004;72(12):7275-81.
      Species: Human
      Sample Type: Nasal Lavage
    24. 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
    25. T lymphocyte-mediated changes in airway smooth muscle responsiveness are attributed to induced autocrine release and actions of IL-5 and IL-1beta.
      Authors: Hakonarson H, Whelan R, Leiter J, Kim C, Chen M, Campbell D, Grunstein MM
      J. Allergy Clin. Immunol., 2002;110(4):624-33.
      Species: Human
      Sample Type: Cell Culture Supernates
    26. Relationship between the perception of dyspnoea and airway inflammatory markers.
      Authors: Jang AS, Choi IS
      Respir Med, 2002;96(3):150-4.
      Species: Human
      Sample Type: Saliva
    27. Structural features of peptide analogs of human histocompatibility leukocyte antigen class I epitopes that are more potent and immunogenic than wild-type peptide.
      Authors: Tangri S, Ishioka GY, Huang X, Sidney J, Southwood S, Fikes J, Sette A
      J. Exp. Med., 2001;194(6):833-46.
      Species: Human
      Sample Type: Cell Culture Supernates
    28. Airway inflammation in asthma and perennial allergic rhinitis. Relationship with nonspecific bronchial responsiveness and maximal airway narrowing.
      Authors: Alvarez MJ, Olaguibel JM, Garcia BE, Rodriquez A, Tabar AI, Urbiola E
      Allergy, 2000;55(4):355-62.
      Species: Human
      Sample Type: Sputum
    29. Low levels of interferon-gamma in nasal fluid accompany raised levels of T-helper 2 cytokines in children with ongoing allergic rhinitis.
      Authors: Benson M, Strannegard IL, Wennergren G, Strannegard O
      Pediatr Allergy Immunol, 2000;11(1):20-8.
      Species: Human
      Sample Type: Nasal Lavage
    30. Bleomycin stimulates lung fibroblast and epithelial cell lines to release eosinophil chemotactic activity.
      Authors: Sato E, Koyama S
      Eur. Respir. J., 2000;16(5):951-8.
      Species: Human
      Sample Type: Cell Culture Supernates
    31. Natural interferon alpha/beta-producing cells link innate and adaptive immunity.
      Authors: Kadowaki N, Antonenko S, Lau JY, Liu YJ
      J. Exp. Med., 2000;192(2):219-26.
      Species: Human
      Sample Type: Cell Culture Supernates
    Expand to show all 31 Citations
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