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Human IL-4 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
31.2 - 2,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
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 IL-4. The suggested diluent is suitable for the analysis of most cell culture supernate 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 IL-4 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
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-4

IL-4 (Interleukin-4) is a cytokine that is primarily expressed by Th2-biased CD4+ T cells, mast cells, basophils, and eosinophils. It promotes cell proliferation, survival, and immunoglobulin class switch to IgG4 and IgE in human B cells, acquisition of the Th2 phenotype by naïve CD4+ T cells, priming and chemotaxis of mast cells, eosinophils, and basophils, and the proliferation and activation of epithelial cells. IL-4 plays a dominant role in the development of allergic inflammation and asthma. The type I receptor for IL-4, which is expressed on hematopoietic cells, is a heterodimer of the ligand binding IL-4 R? and the common ? chain (a shared subunit of the receptors for IL-2, -7, -9, -15, and -21). The type II receptor on nonhematopoietic cells consists of IL-4 R? and IL-13 R?1. The type II receptor also transduces IL-13 mediated signals.

Long Name:
Interleukin 4
Entrez Gene IDs:
3565 (Human); 16189 (Mouse); 287287 (Rat); 397225 (Porcine); 280824 (Bovine); 403785 (Canine); 574281 (Primate); 100302454 (Rabbit)
Alternate Names:
B cell growth factor 1; BCDF; B-cell stimulatory factor 1; BCGF1; BCGF-1; binetrakin; BSF1; BSF-1; IL4; IL-4; IL-4B_cell stimulatory factor 1; IL4E12; interleukin 4; interleukin-4; Lymphocyte stimulatory factor 1; MGC79402; pitrakinra

Assay Procedure

GENERAL ELISA PROTOCOL

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-4 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.

37 Citations: Showing 1 - 10
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  1. Cytokine Profile and Anti-Inflammatory Activity of a Standardized Conditioned Medium Obtained by Coculture of Monocytes and Mesenchymal Stromal Cells (PRS CK STORM)
    Authors: JP Lapuente, A Blázquez-M, J Marco-Brua, G Gómez, P Desportes, J Sanz, P Fernández, M García-Gil, F Bermejo, JV San Martín, A Algaba, JC De Gregori, D Lapuente, A De Gregori, B Lapuente, MV Andrés, A Anel
    Biomolecules, 2022;12(4):.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Reduced Interleukin-17-Expressing Cells in Cutaneous Melanoma
    Authors: A Tosi, L Nardinocch, ML Carbone, L Capriotti, E Pagani, S Mastroeni, C Fortes, F Scopelliti, C Cattani, F Passarelli, A Rosato, S D'Atri, CM Failla, A Cavani
    Biomedicines, 2021;9(12):.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Neutrophils promote T-cell activation through the regulated release of CD44-bound Galectin-9 from the cell surface during HIV infection
    Authors: G Dunsmore, EP Rosero, S Shahbaz, DM Santer, J Jovel, P Lacy, S Houston, S Elahi
    PloS Biology, 2021;19(8):e3001387.
    Species: Human
    Sample Types: Plasma
  4. Altered plasma levels of &betaC and &gammaC chain cytokines and post-treatment modulation in tuberculous lymphadenitis
    Authors: GR Kathamuthu, K Moideen, R Sridhar, D Baskaran, S Babu
    Cytokine, 2021;138(0):155405.
    Species: Human
    Sample Types: Plasma
  5. New Biomarker in Chagas Disease: Extracellular Vesicles Isolated from Peripheral Blood in Chronic Chagas Disease Patients Modulate the Human Immune Response
    Authors: RP Madeira, LM Dal'Mas Ro, P de Cássia, C Mady, BM Ianni, AC Torrecilha
    Journal of Immunology Research, 2021;2021(0):6650670.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Impact of the Uncoupling Protein 1 on Cardiovascular Risk in Patients with Rheumatoid Arthritis
    Authors: LI Lyngfelt, MC Erlandsson, M Nadali, S Hedjazifar, R Pullerits, KM Andersson, P Brembeck, ST Silfverswä, U Smith, MI Bokarewa
    Cells, 2021;10(5):.
    Species: Human
    Sample Types: Serum
  7. Altered plasma levels of &betaC and &gammaC chain cytokines and post-treatment modulation in tuberculous lymphadenitis
    Authors: GR Kathamuthu, K Moideen, R Sridhar, D Baskaran, S Babu
    Cytokine, 2020;138(0):155405.
    Species: Human
    Sample Types: Plasma
  8. Monocytes differentiated into macrophages and dendritic cells in the presence of human IFN-&lambda3 or IFN-&lambda4 show distinct phenotypes
    Authors: M De, A Bhushan, S Chinnaswam
    J Leukoc Biol, 2020;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Circulating growth/differentiation factor 15 is associated with human CD56bright natural killer cell dysfunction and nosocomial infection in severe systemic inflammation
    Authors: H Kleinertz, M Hepner-Sch, S Ehnert, M Claus, R Halbgebaue, L Boller, M Huber-Lang, P Cinelli, C Kirschning, S Flohé, A Sander, C Waydhas, S Vonderhage, M Jäger, M Dudda, C Watzl, SB Flohé
    EBioMedicine, 2019;0(0):.
    Species: Human
    Sample Types: Serum
  10. In Vitro Induction of T Helper 17 Cells by Synergistic Activation of Human Monocyte-Derived Langerhans Cell-Like Cells with Bacterial Agonists
    Authors: R Gramlich, E Aliahmadi, M Peiser
    Int J Mol Sci, 2019;20(6):.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Large-scale secretome analyses unveil the superior immunosuppressive phenotype of umbilical cord stromal cells as compared to other adult mesenchymal stromal cells
    Authors: A Islam, I Urbarova, JA Bruun, I Martinez-Z
    Eur Cell Mater, 2019;37(0):153-174.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Annexin A1, FPR2/ALX, and inflammatory cytokine expression in peritoneal endometriosis
    Authors: LK Volpato, VV Horewicz, F Bobinski, DF Martins, AP Piovezan
    J. Reprod. Immunol., 2018;129(0):30-35.
    Species: Human
    Sample Types: Peritoneal Fluid
  13. An Autocrine Cytokine/JAK/STAT-Signaling Induces Kynurenine Synthesis in Multidrug Resistant Human Cancer Cells.
    Authors: Campia I, Buondonno I, Castella B, Rolando B, Kopecka J, Gazzano E, Ghigo D, Riganti C
    PLoS ONE, 2015;10(5):e0126159.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Characterization of host and microbial determinants in individuals with latent tuberculosis infection using a human granuloma model.
    Authors: Guirado E, Mbawuike U, Keiser T, Arcos J, Azad A, Wang S, Schlesinger L
    MBio, 2015;6(1):e02537-14.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Human CD4+ T cell responses to the dog major allergen Can f 1 and its human homologue tear lipocalin resemble each other.
    Authors: Liukko A, Kinnunen T, Rytkonen-Nissinen M, Kailaanmaki A, Randell J, Maillere B, Virtanen T
    PLoS ONE, 2014;9(5):e98461.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Elevated adaptive immune responses are associated with latent infections of Wuchereria bancrofti.
    Authors: Arndts K, Deininger S, Specht S, Klarmann U, Mand S, Adjobimey T, Debrah AY, Batsa L, Kwarteng A, Epp C, Taylor M, Adjei O, Layland LE, Hoerauf A
    PLoS Negl Trop Dis, 2012;6(4):e1611.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Possible involvement of IL-21 and IL-10 on salivary IgA levels in chronic periodontitis subjects.
    Authors: Napimoga MH, Nunes LH, Maciel AA, Demasi AP, Benatti BB, Santos VR, Bastos MF, de Miranda TS, Duarte PM
    Scand. J. Immunol., 2011;74(6):596-602.
    Species: Human
    Sample Types: Tissue Homogenates
  18. IL-17 amplifies human contact hypersensitivity by licensing hapten nonspecific Th1 cells to kill autologous keratinocytes.
    Authors: Pennino D, Eyerich K, Scarponi C, Carbone T, Eyerich S, Nasorri F, Garcovich S, Traidl-Hoffmann C, Albanesi C, Cavani A
    J. Immunol., 2010;184(9):4880-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Immunoregulation of autocrine prolactin: suppressing the expression of costimulatory molecules and cytokines in T lymphocytes by prolactin receptor knockdown.
    Authors: Xu D, Lin L, Lin X
    Cell. Immunol., 2010;263(1):71-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. TLR2-activated human langerhans cells promote Th17 polarization via IL-1beta, TGF-beta and IL-23.
    Authors: Aliahmadi E, Gramlich R, Grutzkau A, Hitzler M, Kruger M, Baumgrass R, Schreiner M, Wittig B, Wanner R, Peiser M
    Eur. J. Immunol., 2009;39(5):1221-30.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. Der p 1 suppresses indoleamine 2, 3-dioxygenase in dendritic cells from house dust mite-sensitive patients with asthma.
    Authors: Maneechotesuwan K, Wamanuttajinda V, Kasetsinsombat K, Huabprasert S, Yaikwawong M, Barnes PJ, Wongkajornsilp A
    J. Allergy Clin. Immunol., 2009;123(1):239-48.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Interferon-beta treatment in multiple sclerosis attenuates inflammatory gene expression through inducible activity of the phosphatase SHP-1.
    Authors: Christophi GP, Panos M, Hudson CA, Tsikkou C, Mihai C, Mejico LJ, Jubelt B, Massa PT
    Clin. Immunol., 2009;133(1):27-44.
    Species: Human
    Sample Types: Plasma
  23. Fungal proteases induce Th2 polarization through limited dendritic cell maturation and reduced production of IL-12.
    Authors: Lamhamedi-Cherradi SE, Martin RE, Ito T, Kheradmand F, Corry DB, Liu YJ, Moyle M
    J. Immunol., 2008;180(9):6000-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. Helminth infections associated with multiple sclerosis induce regulatory B cells.
    Authors: Correale J, Farez M, Razzitte G
    Ann. Neurol., 2008;64(2):187-99.
    Species: Human
    Sample Types: Cell Culture Supernates
  25. Neuroimmune crosstalk in asthma: dual role of the neurotrophin receptor p75NTR.
    Authors: Nassenstein C, Kammertoens T, Veres TZ, Uckert W, Spies E, Fuchs B, Krug N, Braun A
    J. Allergy Clin. Immunol., 2007;120(5):1089-96.
    Species: Mouse
    Sample Types: BALF
  26. Human epithelial cells establish direct antifungal defense through TLR4-mediated signaling.
    Authors: Weindl G, Naglik JR, Kaesler S, Biedermann T, Hube B, Korting HC, Schaller M
    J. Clin. Invest., 2007;117(12):3664-3672.
    Species: Human
    Sample Types: Cell Culture Supernates
  27. Tumor-associated leukemia inhibitory factor and IL-6 skew monocyte differentiation into tumor-associated macrophage-like cells.
    Authors: Duluc D, Delneste Y, Tan F, Moles MP, Grimaud L, Lenoir J, Preisser L, Anegon I, Catala L, Ifrah N, Descamps P, Gamelin E, Gascan H, Hebbar M, Jeannin P
    Blood, 2007;110(13):4319-30.
    Species: Human
    Sample Types: Cell Culture Supernates
  28. Respiratory syncytial virus synergizes with Th2 cytokines to induce optimal levels of TARC/CCL17.
    Authors: Monick MM, Powers LS, Hassan I, Groskreutz D, Yarovinsky TO, Barrett CW, Castilow EM, Tifrea D, Varga SM, Hunninghake GW
    J. Immunol., 2007;179(3):1648-58.
    Species: Human
    Sample Types: Cell Culture Supernates
  29. An investigation of auto-reactivity after head injury.
    Authors: Cox AL, Coles AJ, Nortje J, Bradley PG, Chatfield DA, Thompson SJ, Menon DK
    J. Neuroimmunol., 2006;174(1):180-6.
    Species: Human
    Sample Types: Cell Culture Supernates
  30. Mast cells regulate procollagen I (alpha 1) production by bronchial fibroblasts derived from subjects with asthma through IL-4/IL-4 delta 2 ratio.
    Authors: Plante S, Semlali A, Joubert P, Bissonnette E, Laviolette M, Hamid Q, Chakir J
    J. Allergy Clin. Immunol., 2006;117(6):1321-7.
    Species: Human
    Sample Types: Cell Culture Supernates
  31. Manipulation of host cytokine network by ticks: a potential gateway for pathogen transmission.
    Authors: Hajnicka V, Vancova I, Kocakova P, Slovak M, Gasperik J, Slavikova M, Hails RS, Labuda M, Nuttall PA
    Parasitology, 2005;130(0):333-42.
    Species: Human
    Sample Types: Saliva
  32. Microbial exposure, symptoms and inflammatory mediators in nasal lavage fluid of kitchen and clerical personnel in schools.
    Authors: Lignell U, Meklin T, Putus T, Vepsalainen A, Roponen M, Torvinen E, Reeslev M, Pennanen S, Hirvonen MR, Kalliokoski P, Nevalainen A
    Int J Occup Med Environ Health, 2005;18(2):139-50.
    Species: Human
    Sample Types: Nasal Lavage Fluid
  33. Immune responses in normal Indian langur monkeys (Presbytis entellus)--a primate model for visceral leishmaniasis.
    Authors: Misra A, Dube A, Naik S
    J. Med. Primatol., 2004;33(2):65-9.
    Species: Primate - Presbytis entellus (Indian langur)
    Sample Types: Cell Culture Supernates
  34. The Toll-like receptor-2/6 agonist macrophage-activating lipopeptide-2 cooperates with IFN-gamma to reverse the Th2 skew in an in vitro allergy model.
    Authors: Weigt H, Muhlradt PF, Larbig M, Krug N, Braun A
    J. Immunol., 2004;172(10):6080-6.
    Species: Human
    Sample Types: Cell Culture Supernates
  35. Comparison of inflammatory elements in nasal lavage and induced sputum following occupational exposure to moldy-building microbes.
    Authors: Purokivi M, Hirvonen MR, Roponen M, Randell J, Vahteristo M, Tukiainen H
    Inhal Toxicol, 2002;14(6):653-62.
    Species: Human
    Sample Types: Sputum
  36. Inflammatory mediators in nasal lavage, induced sputum and serum of employees with rheumatic and respiratory disorders.
    Authors: Roponen M, Kiviranta J, Seuri M, Tukiainen H, Myllykangas-Luosujarvi R, Hirvonen MR
    Eur. Respir. J., 2001;18(3):542-8.
    Species: Human
    Sample Types: Serum
  37. Reproducibility of measurements of exhaled NO, and cell count and cytokine concentrations in induced sputum.
    Authors: Purokivi M, Randell J, Hirvonen MR, Tukiainen H
    Eur. Respir. J., 2000;16(2):242-6.
    Species: Human
    Sample Types: Sputum

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Reviews for Human IL-4 DuoSet ELISA

Average Rating: 4.8 (Based on 4 Reviews)

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Human IL-4 DuoSet ELISA
By Anonymous on 10/09/2020
Sample Tested: Cell culture supernatant

Human IL-4 DuoSet ELISA
By Tahmineh Safaie on 06/16/2020
Sample Tested: Blood mononuclear cells (PBMCs)

Human IL-4 DuoSet ELISA
By Anonymous on 12/16/2019
Sample Tested: primary T cells

Human IL-4 DuoSet ELISA
By Maria Yanez on 05/10/2018
Sample Tested: M2 macrophage