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Human IFN-gamma ELISpot Kit

  • 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.


PRODUCT SUMMARY
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.


PRODUCT FEATURES

  • 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


KIT CONTENTS

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

OTHER REAGENTS REQUIRED

  • 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
  • Long Name:
    Interferon gamma
  • Aliases:
    IFNG; IFG; IFI; IFN-gamma; Immune interferon; interferon gamma; interferon, gamma
  • Entrez Gene IDs:
    3458 (Human); 15978 (Human); 396991 (Human); 281237 (Human); 25712 (Rat); 403801 (Canine); 493965 (Feline);
  • Background:
    IFN-gamma

Interferon-gamma (IFN-gamma, also known as Type II interferon or immune interferon) is a cytokine produced primarily by T-lymphocytes and natural killer cells. The protein shares no significant homology with IFN-beta or the various IFN-alpha family proteins. Mature IFN-gamma exists as noncovalently-linked homodimers. Human IFN-gamma is highly species specific and is biologically active only in human and primate cells.

IFN-gamma was originally characterized based on its antiviral activities. The protein also exerts anti-proliferative, immunoregulatory and proinflammatory activities and is thus important in host defense mechanisms. IFN-gamma induces the production of cytokines, up-regulates the expression of class I and II MHC antigens, Fc receptor and leukocyte adhesion molecules. It modulates macrophage effector functions, influences isotype switching and potentiates the secretion of immunoglobulins by B cells. IFN-gamma also augments Helper T cell expansion and may be required for Helper T cell differentiation.

IFN-gamma exerts its biological activities by binding to specific cell surface receptors with high-affinity binding sites. The IFN-gamma receptor is present on almost all cell types except mature erythrocytes and has been cloned and characterized. The IFN-gamma receptor is structurally related to the IL-10 receptor.

Citations:

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.

Showing Results 1 - 10 of 13
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Species
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
  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)
  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
  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.
  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
  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.
  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
  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
  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
  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
  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
  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.
  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
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