Human IL-2 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY202
DY202-05
Ancillary Products Available
Human IL-2 ELISA Standard Curve
1 Image
Product Details
Procedure
Citations (29)
FAQs
Supplemental Products
Reviews (2)

Human IL-2 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
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-2. 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)

duct.

 

Data Example

Human IL-2 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: IL-2

IL-2 (Interleukin 2) is a critical cytokine in T cell biology. It promotes T cell activation and expansion, the development, maintenance and function of regulatory T cells (Treg), and the differentiation of CD8+ T cells into terminal effector cells and memory cells. IL-2 signals through a receptor complex composed of CD25/IL-2 R alpha, IL-2 R beta, and the Common gamma Chain (gamma c). IL-2 R beta is also a component of the IL-15 receptor complex. Gamma c is also a signaling subunit in the receptors for IL-4, -7, -9, -15, and -21.

Long Name:
Interleukin 2
Entrez Gene IDs:
3558 (Human); 16183 (Mouse); 116562 (Rat); 396868 (Porcine); 280822 (Bovine); 403989 (Canine); 100034204 (Equine); 751114 (Feline); 100302458 (Rabbit)
Alternate Names:
aldesleukin; IL2; IL-2; IL-2lymphokine; interleukin 2; interleukin-2; involved in regulation of T-cell clonal expansion; T cell growth factor; T-cell growth factor; TCGF

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-2 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.

29 Citations: Showing 1 - 10
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  1. Interferon-? converts human microvascular pericytes into negative regulators of alloimmunity through induction of indoleamine 2,3-dioxygenase 1
    Authors: R Liu, J Merola, TD Manes, L Qin, GT Tietjen, F López-Girá, V Broecker, C Fang, C Xie, PM Chen, NC Kirkiles-S, D Jane-Wit, JS Pober
    JCI Insight, 2018;3(5):.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Identification of benzazole compounds that induce HIV-1 transcription
    Authors: JD Graci, D Michaels, G Chen, GM Schiralli, S Nodder, M Weetall, GM Karp, Z Gu, JM Colacino, AJ Henderson
    PLoS ONE, 2017;12(6):e0179100.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. One-domain CD4 Fused to Human Anti-CD16 Antibody Domain Mediates Effective Killing of HIV-1-Infected Cells
    Authors: W Li, Y Wu, D Kong, H Yang, Y Wang, J Shao, Y Feng, W Chen, L Ma, T Ying, DS Dimitrov
    Sci Rep, 2017;7(1):9130.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Chemical proteomic map of dimethyl fumarate-sensitive cysteines in primary human T cells
    Authors: Benjamin F Cravatt
    Sci Signal, 2016;9(445):rs10.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  5. Additive anti-inflammatory effects of corticosteroids and phosphodiesterase-4 inhibitors in COPD CD8 cells.
    Authors: Grundy S, Plumb J, Kaur M, Ray D, Singh D
    Respir Res, 2016;17(1):9.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Rac1-Rab11-FIP3 regulatory hub coordinates vesicle traffic with actin remodeling and T-cell activation
    EMBO J, 2016;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Reduced systemic and mycobacterial antigen-stimulated concentrations of IL-1? and IL-18 in tuberculous lymphadenitis
    Cytokine, 2016;90(0):66-72.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. 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
  9. The tuberculosis vaccine candidate Bacillus Calmette-Guerin DeltaureC::hly coexpressing human interleukin-7 or -18 enhances antigen-specific T cell responses in mice.
    Authors: Rao M, Vogelzang A, Kaiser P, Schuerer S, Kaufmann S, Gengenbacher M
    PLoS ONE, 2015;8(11):e78966.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Engagement of SLAMF2/CD48 prolongs the time frame of effective T cell activation by supporting mature dendritic cell survival.
    Authors: Kis-Toth K, Tsokos G
    J Immunol, 2014;192(9):4436-42.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Anti-inflammatory effects of green soybean extract irradiated with visible light.
    Authors: Tanaka K, Ohgo Y, Katayanagi Y, Yasui K, Hiramoto S, Ikemoto H, Nakata Y, Miyoshi N, Isemura M, Ohashi N, Imai S
    Sci Rep, 2014;4(0):4732.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. 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;44(5):1330-40.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. T-cell engager-armed oncolytic vaccinia virus significantly enhances antitumor therapy.
    Authors: Yu F, Wang X, Guo Z, Bartlett D, Gottschalk S, Song X
    Mol Ther, 2014;22(1):102-11.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Dynamic motile T cells highly respond to the T cell stimulation via PI3K-Akt and NF-kappaB pathways.
    Authors: Kim H, Na B, Kwon M, Ko Y, Han W, Jun C
    PLoS ONE, 2013;8(3):e59793.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Leukocyte function-associated antigen-1/intercellular adhesion molecule-1 interaction induces a novel genetic signature resulting in T-cells refractory to transforming growth factor-beta signaling.
    Authors: Verma N, Dempsey E, Long A, Davies A, Barry S, Fallon P, Volkov Y, Kelleher D
    J Biol Chem, 2012;287(32):27204-16.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. 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
  17. IL-8 dictates glycosaminoglycan binding and stability of IL-18 in cystic fibrosis.
    Authors: Reeves EP, Williamson M, Byrne B, Bergin DA, Smith SG, Greally P, O'Kennedy R, O'Neill SJ, McElvaney NG
    J. Immunol., 2010;184(3):1642-52.
    Species: Human
    Sample Types: Serum
  18. 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
  19. 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
  20. Calcium dependence of T cell proliferation following focal stimulation.
    Authors: Schwarz EC, Kummerow C, Wenning AS, Wagner K, Sappok A, Waggershauser K, Griesemer D, Strauss B, Wolfs MJ, Quintana A, Hoth M
    Eur. J. Immunol., 2007;37(10):2723-33.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. Immunotherapy for neuroblastoma using syngeneic fibroblasts transfected with IL-2 and IL-12.
    Authors: Barker SE, Grosse SM, Siapati EK, Kritz A, Kinnon C, Thrasher AJ, Hart SL
    Br. J. Cancer, 2007;97(2):210-7.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Familial NK cell deficiency associated with impaired IL-2- and IL-15-dependent survival of lymphocytes.
    Authors: Eidenschenk C, Jouanguy E, Alcais A, Mention JJ, Pasquier B, Fleckenstein IM, Puel A, Gineau L, Carel JC, Vivier E, Le Deist F, Casanova JL
    J. Immunol., 2006;177(12):8835-43.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. The Wingless homolog WNT5A and its receptor Frizzled-5 regulate inflammatory responses of human mononuclear cells induced by microbial stimulation.
    Authors: Blumenthal A, Ehlers S, Lauber J, Buer J, Lange C, Goldmann T, Heine H, Brandt E, Reiling N
    Blood, 2006;108(3):965-73.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. Splice variants of human FOXP3 are functional inhibitors of human CD4+ T-cell activation.
    Authors: Smith EL, Finney HM, Nesbitt AM, Ramsdell F, Robinson MK
    Immunology, 2006;119(2):203-11.
    Species: Human
    Sample Types: Cell Culture Supernates
  25. 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
  26. Human T cells constitutively express IL-15 that promotes ex vivo T cell homeostatic proliferation through autocrine/juxtacrine loops.
    Authors: Miranda-Carus ME, Benito-Miguel M, Llamas MA, Balsa A, Martin-Mola E
    J. Immunol., 2005;175(6):3656-62.
    Species: Human
    Sample Types: Cell Culture Supernates
  27. Activation of resting human primary T cells with chimeric receptors: costimulation from CD28, inducible costimulator, CD134, and CD137 in series with signals from the TCR zeta chain.
    Authors: Finney HM, Akbar AN, Lawson AD
    J. Immunol., 2004;172(1):104-13.
    Species: Human
    Sample Types: Cell Culture Supernates
  28. 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
  29. Mycophenolic acid inhibits IL-2-dependent T cell proliferation, but not IL-2-dependent survival and sensitization to apoptosis.
    Authors: Quemeneur L, Flacher M, Gerland LM, Ffrench M, Revillard JP, Bonnefoy-Berard N
    J. Immunol., 2002;169(5):2747-55.
    Species: Human
    Sample Types: Cell Culture Supernates

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Reviews for Human IL-2 DuoSet ELISA

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Human IL-2 DuoSet ELISA
By Anonymous on 08/06/2018
Sample Tested: Serum

Human IL-2 DuoSet ELISA
By Anonymous on 10/20/2017
Sample Tested: Human cells