Human IL-5 DuoSet ELISA

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Catalog # Availability Size / Price Qty
DY205
DY205-05
Ancillary Products Available

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Human IL-5 ELISA Standard Curve
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Product Details
Procedure
Citations (22)
FAQs
Supplemental Products
Reviews (3)

Human IL-5 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
4.7 - 300 pg/mL
Sufficient Materials
For five or 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-5. 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 ELISA.

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

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, 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)

Scientific Data

Human IL-5 ELISA Standard Curve

Product Datasheets

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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-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

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 Reagent Diluent 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-5 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.

22 Citations: Showing 1 - 10
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  1. Macrophages derived from LPS-stimulated monocytes from individuals with subclinical atherosclerosis were characterized by increased pro-inflammatory activity
    Authors: Nikiforov, NG;Kirichenko, TV;Kubekina, MV;Chegodaev, YS;Zhuravlev, AD;Ilchuk, LA;Nikolaeva, MA;Arefieva, AS;Popov, MA;Verkhova, SS;Bagheri Ekta, M;Orekhov, AN;
    Cytokine
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Inflammatory biomarkers at different stages of Sarcopenia in older women
    Authors: da Costa Teixeira, LA;Avelar, NCP;Peixoto, MFD;Parentoni, AN;Santos, JMD;Pereira, FSM;Danielewicz, AL;Leopoldino, AAO;Costa, SP;Arrieiro, AN;Soares, LA;da Silva Lage, VK;Prates, ACN;Taiar, R;de Carvalho Bastone, A;Oliveira, VC;Oliveira, MX;Costa, HS;Nobre, JNP;Brant, FP;Duarte, TC;Figueiredo, PHS;Mendonça, VA;Lacerda, ACR;
    Scientific reports
    Species: Human
    Sample Types: Plasma
  3. Adiponectin Is a Contributing Factor of Low Appendicular Lean Mass in Older Community-Dwelling Women: A Cross-Sectional Study
    Authors: LAC Teixeira, JM Dos Santos, AN Parentoni, LP Lima, TC Duarte, FP Brant, CDC Neves, FSM Pereira, NCP Avelar, AL Danielewic, AAO Leopoldino, SP Costa, AN Arrieiro, LA Soares, ACN Prates, JNP Nobre, A de Carvalh, VC de Oliveir, MX Oliveira, PH Scheidt Fi, HS Costa, V Amaral Men, R Taiar, AC Rodrigues
    Journal of Clinical Medicine, 2022-12-02;11(23):.
    Species: Human
    Sample Types: Plasma
  4. Substrate stiffness engineered to replicate disease conditions influence senescence and fibrotic responses in primary lung fibroblasts
    Authors: KEC Blokland, M Nizamoglu, H Habibie, T Borghuis, M Schuliga, BN Melgert, DA Knight, CA Brandsma, SD Pouwels, JK Burgess
    Frontiers in Pharmacology, 2022-11-03;13(0):989169.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Anti-inflammatory effects of an autologous gold-based serum therapy in osteoarthritis patients
    Authors: J Feldt, AJ Donaubauer, J Welss, U Schneider, US Gaipl, F Paulsen
    Scientific Reports, 2022-03-03;12(1):3560.
    Species: Human
    Sample Types: Serum
  6. Bacterial genotoxins induce T�cell senescence
    Authors: SL Mathiasen, L Gall-Mas, IS Pateras, SDP Theodorou, MRJ Namini, MB Hansen, OCB Martin, CK Vadivel, K Ntostoglou, D Butter, M Givskov, C Geisler, AN Akbar, VG Gorgoulis, T Frisan, N Ødum, T Krejsgaard
    Cell Reports, 2021-06-08;35(10):109220.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Effectiveness and safety of mepolizumab in combination with corticosteroids in patients with eosinophilic granulomatosis with polyangiitis
    Authors: M Ueno, I Miyagawa, K Nakano, S Iwata, K Hanami, S Fukuyo, S Kubo, Y Miyazaki, A Kawabe, H Yoshinari, S Nakayamada, Y Tanaka
    Arthritis Research & Therapy, 2021-03-16;23(1):86.
    Species: Human
    Sample Types: Serum
  8. 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, 2021-01-11;2021(0):6650670.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. 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, 2020-12-17;138(0):155405.
    Species: Human
    Sample Types: Plasma
  10. Salivary IL-8 and IL-1RA changes induced by exercise in cold-dry and normal conditions
    Authors: I Patlan, MP Jeffrey, D Lewis, J Green-John, S Dogra
    Cytokine, 2020-10-28;137(0):155347.
    Species: Human
    Sample Types: Saliva
  11. Altered miR-155 expression in Allergic Asthmatic airways
    Authors: C Malmhäll, K Johansson, C Winkler, S Alawieh, L Ekerljung, M Rådinger
    Scand. J. Immunol, 2017-04-01;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Genetic, immunological, and clinical features of patients with bacterial and fungal infections due to inherited IL-17RA deficiency
    Proc. Natl. Acad. Sci. U.S.A, 2016-12-07;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. 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, 2014-05-29;9(5):e98461.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. 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, 2014-02-20;8(2):e2679.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. The aryl hydrocarbon receptor is functionally upregulated early in the course ofvhuman T-cell activation.
    Authors: Prigent L, Robineau M, Jouneau S, Morzadec C, Louarn L, Vernhet L, Fardel O, Sparfel L
    Eur J Immunol, 2014-02-18;44(5):1330-40.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. An ACT1 mutation selectively abolishes interleukin-17 responses in humans with chronic mucocutaneous candidiasis.
    Authors: Boisson B, Wang C, Pedergnana V, Wu L, Cypowyj S, Rybojad M, Belkadi A, Picard C, Abel L, Fieschi C, Puel A, Li X, Casanova J
    Immunity, 2013-10-10;39(4):676-86.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. 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, 2012-04-03;6(4):e1611.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. Natural killer cell-mediated cytokine response among HIV-positive south Indians with pulmonary tuberculosis.
    Authors: Ramana Rao PV, Rajasekaran S, Raja A
    J. Interferon Cytokine Res., 2010-01-01;30(1):33-42.
    Species: Human
    Sample Types: Plasma
  19. 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., 2008-05-01;180(9):6000-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. 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., 2007-08-22;120(5):1089-96.
    Species: Mouse
    Sample Types: BALF
  21. 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., 2006-03-06;174(1):180-6.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Synthesis and biological evaluation of 1,2,4-triazinylphenylalkylthiazolecarboxylic acid esters as cytokine-inhibiting antedrugs with strong bronchodilating effects in an animal model of asthma.
    Authors: Freyne EJ, Lacrampe JF, Deroose F, Boeckx GM, Willems M, Embrechts W, Coesemans E, Willems JJ, Fortin JM, Ligney Y, Dillen LL, Cools WF, Goossens J, Corens D, De Groot A, Van Wauwe JP
    J. Med. Chem., 2005-03-24;48(6):2167-75.
    Species: Human
    Sample Types: Cell Culture Supernates

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Reviews for Human IL-5 DuoSet ELISA

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Human IL-5 DuoSet ELISA
By Anonymous on 11/07/2020
Sample Tested: Peripheral blood monocytes

Human IL-5 DuoSet ELISA
By Evan Williams on 10/28/2020
Sample Tested: Cell Culture Supernates

easy to use kit. IL-5 detectable in PBMCs treated with house dust mite.


Human IL-5 DuoSet ELISA
By Anonymous on 12/16/2019
Sample Tested: TH-2 CELLS