Human IL-27 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY2526
DY2526-05
Ancillary Products Available
Human IL-27 ELISA Standard Curve
1 Image
Product Details
Procedure
Citations (22)
FAQs
Supplemental Products
Reviews (2)

Human IL-27 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
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-27. 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

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)

 

  

Data Example

Human IL-27 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-27

IL-27 is a heterodimeric cytokine that is composed of EBI3 (which is related to the p40 subunit of IL-12 and IL-23) and p28 (which is related to the p35 chain of IL-12). IL-27 is expressed by monocytes, endothelial cells, and dendritic cells, and it signals through a receptor complex composed of IL-27 R alpha/WSX-1/TCCR and gp130. IL-27 induces IL-12 receptor expression on naïve CD4+ T cells, sensitizing them to subsequent IL-12 induced Th1 development. It also functions as an anti-inflammatory cytokine by limiting T and NKT cell activity.

Long Name:
Interleukin 27
Entrez Gene IDs:
246778 (Human); 246779 (Mouse); 102121242 (Cynomolgus Monkey)
Alternate Names:
IL-27 p28 subunit; IL-27 subunit alpha; IL27; IL-27; IL27A; IL-27-A; IL27-A; IL27p28; IL30; interleukin 27; interleukin 30; interleukin-27 subunit alpha; MGC71873; p28IL-27A

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-27 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. Association between IL-27 and Tr1 cells in severe form of paracoccidioidomycosis
    Authors: L Moreira Ge, L de Oliveir, A da Silva J, L Furquim de, AK Felício Ba, AE Rizzato, P Trabasso, RL Mamoni, R Mendes Per, ML Cintra, L Nunes Sant, M de Carvalh, L Pereira Ru, MH de Souza L
    Cytokine, 2019;127(0):154962.
    Species: Human
    Sample Types: Serum
  2. Heightened systemic levels of anti-inflammatory cytokines in pulmonary tuberculosis and alterations following anti-tuberculosis treatment
    Authors: K Moideen, NP Kumar, R Bethunaick, VV Banurekha, D Nair, S Babu
    Cytokine, 2019;127(0):154929.
    Species: Human
    Sample Types: Plasma
  3. Changes in interleukin-27 levels in patients with acute coronary syndrome and their clinical significance
    Authors: L Zhang, J Zhang, S Su, S Luo
    PeerJ, 2019;7(0):e5652.
    Species: Human
    Sample Types: Plasma
  4. cIAP1/2-TRAF2-SHP-1-Src-MyD88 Complex Regulates Lipopolysaccharide-Induced IL-27 Production through NF-?B Activation in Human Macrophages
    Authors: A Busca, Y Konarski, N Gajanayaka, S O'Hara, J Angel, M Kozlowski, A Kumar
    J. Immunol., 2018;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Influenza infection directly alters innate IL-23 and IL-12p70 and subsequent IL-17A and IFN-? responses to pneumococcus in vitro in human monocytes
    Authors: ST Loughran, PA Power, PT Maguire, SL McQuaid, PJ Buchanan, I Jonsdottir, RW Newman, R Harvey, PA Johnson
    PLoS ONE, 2018;13(9):e0203521.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Human M2 Macrophages Limit NK Cell Effector Functions through Secretion of TGF-? and Engagement of CD85j
    Authors: SY Nuñez, A Ziblat, F Secchiari, NI Torres, JM Sierra, XL Raffo Irao, RE Araya, CI Domaica, MB Fuertes, NW Zwirner
    J. Immunol., 2017;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. In vitro and in vivo antagonistic activity of new probiotic culture against Clostridium difficile and Clostridium perfringens
    Authors: N Goli?, K Veljovi?, N Popovi?, J Djoki?, I Strahini?, I Mrvaljevi?, A Terzi?-Vid
    BMC Microbiol., 2017;17(1):108.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. On the cytokine/chemokine network during Plasmodium vivax malaria: new insights to understand the disease
    Authors: NS Hojo-Souza, DB Pereira, FS de Souza, TA de Oliveir, MS Cardoso, MS Tada, GM Zanini, DC Bartholome, RT Fujiwara, LL Bueno
    Malar. J, 2017;16(1):42.
    Species: Human
    Sample Types: Plasma
  9. Human Immunodeficiency Virus Type-1 Myeloid Derived Suppressor Cells Inhibit Cytomegalovirus Inflammation through Interleukin-27 and B7-H4
    Authors: A Garg, R Trout, SA Spector
    Sci Rep, 2017;7(0):44485.
    Species: Human
    Sample Types: Plasma
  10. Stromal fibroblasts support dendritic cells to maintain IL-23/Th17 responses after exposure to ionizing radiation
    Authors: A Malecka, Q Wang, S Shah, RV Sutavani, I Spendlove, JM Ramage, J Greensmith, HA Franks, MJ Gough, A Saalbach, PM Patel, AM Jackson
    J Leukoc Biol, 2016;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. IL-27 suppresses SKOV3 cells proliferation by enhancing STAT3 and inhibiting the Akt signal pathway
    Mol Immunol, 2016;78(0):155-163.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. IL-27 Suppresses Antimicrobial Activity in Human Leprosy.
    Authors: Teles R, Kelly-Scumpia K, Sarno E, Rea T, Ochoa M, Cheng G, Modlin R
    J Invest Dermatol, 2015;135(10):2410-7.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Antimony resistant Leishmania donovani but not sensitive ones drives greater frequency of potent T-regulatory cells upon interaction with human PBMCs: role of IL-10 and TGF-beta in early immune response.
    Authors: Guha R, Das S, Ghosh J, Sundar S, Dujardin J, Roy S
    PLoS Negl Trop Dis, 2014;8(7):e2995.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. A soluble form of IL-27Ralpha is a natural IL-27 antagonist.
    Authors: Dietrich C, Candon S, Ruemmele F, Devergne O
    J Immunol, 2014;192(11):5382-9.
    Species: Human
    Sample Types: Serum
  15. Differential reconstitution of T cell subsets following immunodepleting treatment with alemtuzumab (anti-CD52 monoclonal antibody) in patients with relapsing-remitting multiple sclerosis.
    Authors: Zhang X, Tao Y, Chopra M, Ahn M, Marcus K, Choudhary N, Zhu H, Markovic-Plese S
    J Immunol, 2013;191(12):5867-74.
    Species: Human
    Sample Types: Serum
  16. Cytokine and chemokine responses to helminth and protozoan parasites and to fungus and mite allergens in neonates, children, adults, and the elderly.
    Authors: Lechner C, Komander K, Hegewald J, Huang X, Gantin R, Soboslay P, Agossou A, Banla M, Kohler C
    Immun Ageing, 2013;10(1):29.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Independent and interdependent immunoregulatory effects of IL-27, IFN-beta, and IL-10 in the suppression of human Th17 cells and murine experimental autoimmune encephalomyelitis.
    Authors: Fitzgerald D, Fonseca-Kelly Z, Cullimore M, Safabakhsh P, Saris C, Zhang G, Rostami A
    J Immunol, 2013;190(7):3225-34.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. Serum levels of Th17-related cytokines in Behcet disease patients after cataract surgery.
    Authors: Jiang S, Liu X, Luo L, Qu B, Huang X, Lin Y, Ye S, Liu Y
    Mol. Vis., 2011;17(0):1425-30.
    Species: Human
    Sample Types: Serum
  19. B cells as a therapeutic target for IFN-beta in relapsing-remitting multiple sclerosis.
    Authors: Ramgolam VS, Sha Y, Marcus KL, Choudhary N, Troiani L, Chopra M, Markovic-Plese S
    J. Immunol., 2011;186(0):4518.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. IL-27 mediates the response to IFN-beta therapy in multiple sclerosis patients by inhibiting Th17 cells.
    Authors: Sweeney CM, Lonergan R, Basdeo SA, Kinsella K, Dungan LS, Higgins SC, Kelly PJ, Costelloe L, Tubridy N, Mills KH, Fletcher JM
    Brain Behav. Immun., 2010;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. Impact of HIV infection, highly active antiretroviral therapy, and hepatitis C coinfection on serum interleukin-27.
    Authors: Guzzo C, Hopman WM, Mat NF
    AIDS, 2010;24(9):1371-4.
    Species: Human
    Sample Types: Serum
  22. Helminth antigens modulate immune responses in cells from multiple sclerosis patients through TLR2-dependent mechanisms.
    Authors: Correale J, Farez M
    J. Immunol., 2009;183(9):5999-6012.
    Species: Human
    Sample Types: Cell Culture Supernates

FAQs

  1. Is the Human IL-27 DuoSet ELISA (Catalog # DY2526) specific to p28 or EBI3 subunit, or does the kit recognize both subunits?

    • The Human IL-27 DuoSet ELISA is designed to detect the IL27 p28 and EBI3 heterodimer, and therefore will recognize both IL-27 subunits. Additional information can be found in the specificity section of the product datasheet.

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Reviews for Human IL-27 DuoSet ELISA

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Human IL-27 DuoSet ELISA
By Anonymous on 05/06/2019
Application: Sample Tested: Cell culture supernatant

Human IL-27 DuoSet ELISA
By Anonymous on 12/11/2017
Application: Sample Tested: Cell culture supernatant