Human IFN-gamma ELISpot Kit

13 citations
  • Assay Type
    Quantitative Sandwich ELISA
  • Assay Format
    96-well PVDF-backed microplate
  • Assay Length
    3 hours 15 mins to 4 hours 30 mins*
  • Sample Type
    Whole Cells
  • 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.

* Following cell stimulation.

ELISpot kits are highly sensitive, microplate-based assays for the detection of cytokine secreting cells. This kit is designed for the detection and enumeration of . Complete ELISpot kits are ready-to-run and require no assay development or refinement.

This ELISpot assay employs a capture antibody specific for , pre-coated onto a PVDF-backed microplate. Appropriately stimulated cells are pipetted directly into the wells and the immobilized antibody in the immediate vicinity of the secreting cells binds secreted . Following wash steps and incubation with a biotinylated detection antibody, alkaline-phosphatase conjugated to streptavidin is added. Unbound enzyme is subsequently removed by washing and a substrate solution (BCIP/NBT) is added. A blue-black colored precipitate forms at the sites of cytokine localization and appears as spots, with each individual spot representing an individual secreting cell. The spots can be counted with an automated ELISpot reader system or manually using a stereomicroscope.


  • Detect and quantitate individual cells secreting
  • High sensitivity - ELISpot assays can measure responses with frequencies well below 1 in 100,000 cells
  • No in vitro expansion of cells required
  • High-throughput - ELISpot assays use only a small number of primary cells


  • Microplate
  • Biotinylated Detection Antibody
  • Streptavidin conjugated to Alkaline Phosphatase
  • Dilution Buffers
  • Wash Buffer Concentrate
  • BCIP/NBT Chromogen
  • Positive Control


  • Pipettes and pipette tips
  • Deionized or distilled water
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • 500 mL graduated cylinder
  • 37 °C CO2 incubator
  • Sterile culture media
  • Dissection microscope or an automated ELISpot reader

Background: IFN-gamma
IFN-gamma (Interferon-gamma) is the prototype proinflammatory cytokine and is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells. It plays a key role in host defense by promoting the development and activation of Th1 cells, chemoattraction and activation of monocytes and macrophages, upregulation of antigen presentation molecules, and immunoglobulin class switching in B cells. It also exhibits antiviral, antiproliferative, and apoptotic effects. In addition, IFN-gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation. IFN-gamma dimers signal through a receptor complex of two IFN-gamma R1 and two IFN-gamma R2 subunits.
  • Long Name:
    Interferon gamma
  • Aliases:
    IFNG; IFG; IFI; IFN-gamma; Immune interferon; interferon gamma; interferon, gamma

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.

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Sample Type
  1. Comparative Multi-Donor Study of IFNgamma Secretion and Expression by Human PBMCs Using ELISPOT Side-by-Side with ELISA and Flow Cytometry Assays.
    Authors: Hagen J, Zimmerman R, Goetz C, Bonnevier J, Houchins J, Reagan K, Kalyuzhny A
    Cells, 2015;4(1):84-95.
    Species: Human
    Sample Type: whole cells
  2. Antibody-mediated depletion of lymphocyte-activation gene-3 (LAG-3(+) )-activated T lymphocytes prevents delayed-type hypersensitivity in non-human primates.
    Authors: Poirier N, Haudebourg T, Brignone C, Dilek N, Hervouet J, Minault D, Coulon F, de Silly RV, Triebel F, Blancho G, Vanhove B
    Clin. Exp. Immunol., 2011;164(2):265-74.
    Species: Primate – Papio anubis (Olive Baboon)
    Sample Type: Whole Cells
  3. Interferon-gamma is induced in human peripheral blood immune cells in vitro by sodium stibogluconate/interleukin-2 and mediates its antitumor activity in vivo.
    Authors: Fan K, Borden E, Yi T
    J. Interferon Cytokine Res., 2009;29(8):451-60.
    Species: Human
    Sample Type: Whole Cells
  4. Human bronchial intraepithelial T cells produce interferon-gamma and stimulate epithelial cells.
    Authors: Hirosako S, Goto E, Fujii K, Tsumori K, Hirata N, Tsumura S, Kamohara H, Kohrogi H
    Clin. Exp. Immunol., 2009;155(2):266-74.
    Species: Human
    Sample Type: Whole Cells
  5. Phase I therapeutic trial of the HIV-1 Tat protein and long term follow-up.
    Authors: Longo O, Tripiciano A, Fiorelli V, Bellino S, Scoglio A, Collacchi B, Alvarez MJ, Francavilla V, Arancio A, Paniccia G, Lazzarin A, Tambussi G, Din CT, Visintini R, Narciso P, Antinori A, D'Offizi G, Giulianelli M, Carta M, Di Carlo A, Palamara G, Giuliani M, Laguardia ME, Monini P, Magnani M, Ensoli F, Ensoli B
    Vaccine, 2009;27(25):3306-12.
    Species: Human
    Sample Type: Whole Cells
  6. Epitope specificity and clonality of EBV-specific CTLs used to treat posttransplant lymphoproliferative disease.
    Authors: McAulay KA, Haque T, Urquhart G, Bellamy C, Guiretti D, Crawford DH
    J. Immunol., 2009;182(6):3892-901.
    Species: Human
    Sample Type: Whole Cells
  7. Regulatory NK cells suppress antigen-specific T cell responses.
    Authors: Deniz G, Erten G, Kucuksezer UC, Kocacik D, Karagiannidis C, Aktas E, Akdis CA, Akdis M
    J. Immunol., 2008;180(2):850-7.
    Species: Human
    Sample Type: Whole Cells
  8. A multidonor ELISPOT study of IL-1 beta, IL-2, IL-4, IL-6, IL-13, IFN-gamma and TNF-alpha release by cryopreserved human peripheral blood mononuclear cells.
    Authors: Bailey T, Stark S, Grant A, Hartnett C, Tsang M, Kalyuzhny A
    J. Immunol. Methods, 2002;270(2):171-82.
    Species: Human
    Sample Type: Whole Cells
  9. Correlations between vaccinia-specific immune responses within a cohort of armed forces members.
    Authors: Umlauf B, Ovsyannikova I, Haralambieva I, Kennedy R, Vierkant R, Pankratz V, Jacobson R, Poland G
    Viral Immunol, 2011;24(5):415-20.
    Species: Human
    Sample Type: whole cells
  10. Mitoxantrone as rescue therapy in worsening relapsing-remitting MS patients receiving IFN-beta.
    Authors: Correale J, Rush C, Amengual A, Goicochea MT
    J. Neuroimmunol., 2005;162(1):173-83.
    Species: Human
    Sample Type: Whole Cells
  11. Contactin-2/TAG-1-directed autoimmunity is identified in multiple sclerosis patients and mediates gray matter pathology in animals.
    Authors: Derfuss T, Parikh K, Velhin S, Braun M, Mathey E, Krumbholz M, Kumpfel T, Moldenhauer A, Rader C, Sonderegger P, Pollmann W, Tiefenthaller C, Bauer J, Lassmann H, Wekerle H, Karagogeos D, Hohlfeld R, Linington C, Meinl E
    Proc. Natl. Acad. Sci. U.S.A., 2009;106(20):8302-7.
    Species: Human
    Sample Type: Whole Cells
  12. 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 Type: Whole Cells
  13. Impaired hypothalamic-pituitary-adrenal axis activity in patients with multiple sclerosis.
    Authors: Ysrraelit MC, Gaitan MI, Lopez AS, Correale J
    Neurology, 2008;71(24):1948-54.
    Species: Human
    Sample Type: Whole Cells
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