Human IFN-gamma ELISpot Kit

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XEL285
PEL285
EL285
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Product Details
Citations (22)
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Human IFN-gamma ELISpot Kit Summary

Assay Type
Quantitative Sandwich ELISA
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.


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 human IFN-gamma. Complete ELISpot kits are ready-to-run and require no assay development or refinement.

This ELISpot assay employs a capture antibody specific for human IFN-gamma, 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 human IFN-gamma. 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 human IFN-gamma 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 human IFN-gamma
  • 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

  • Human IFN-gamma Microplate
  • Biotinylated Detection Antibody
  • Streptavidin conjugated to Alkaline Phosphatase
  • Dilution Buffers
  • Wash Buffer Concentrate
  • BCIP/NBT Chromogen
  • Human IFN-gamma 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

Product Datasheets

Preparation and Storage

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

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
Entrez Gene IDs:
3458 (Human); 15978 (Mouse); 25712 (Rat); 396991 (Porcine); 281237 (Bovine); 403801 (Canine); 493965 (Feline)
Alternate Names:
IFG; IFI; IFNG; IFNgamma; IFN-gamma; Immune interferon; interferon gamma; interferon, gamma
⚠ WARNING: This product can expose you to chemicals including methanol, which is known to the State of California to cause reproductive toxicity with developmental effects. For more information, go to www.P65Warnings.ca.gov.

Citations for Human IFN-gamma ELISpot Kit

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. Contribution of endotoxin to Th17 bias in patients with non-alcoholic steatohepatitis
    Authors: X Wang, D Ji, B Zhu, S Jiang, L Han, Y Wang, H Mai, S Xu, H Jiang, G Wang, Y Rong
    Microb. Pathog., 2020;0(0):104009.
    Species: Human
    Sample Types: Whole Cells
  2. The first clinical use of a recombinant Lactococcus lactis expressing human papillomavirus type 16 E7 oncogene oral vaccine: A phase I safety and immunogenicity trial in healthy women volunteers
    Authors: AH Mohseni, S Taghinezha, H Keyvani
    Mol. Cancer Ther., 2019;0(0):.
    Species: Human
    Sample Types: Whole Cells
  3. Phase 1 Safety and Immunogenicity Trial of Recombinant Lactococcus lactis Expressing Human Papillomavirus Type 16 E6 Oncoprotein Vaccine
    Authors: S Taghinezha, AH Mohseni, H Keyvani, MR Razavi
    Mol Ther Methods Clin Dev, 2019;15(0):40-51.
    Species: Human
    Sample Types: Whole Cells
  4. Sensitive Detection and Analysis of Neoantigen-Specific T Cell Populations from Tumors and Blood
    Authors: S Peng, JM Zaretsky, AHC Ng, W Chour, MT Bethune, J Choi, A Hsu, E Holman, X Ding, K Guo, J Kim, AM Xu, JE Heath, WJ Noh, J Zhou, Y Su, Y Lu, J McLaughlin, D Cheng, ON Witte, D Baltimore, A Ribas, JR Heath
    Cell Rep, 2019;28(10):2728-2738.e7.
    Species: Human
    Sample Types: Whole Cells
  5. A Potential Antitumor Effect of Dendritic Cells Fused with Cancer Stem Cells in Hepatocellular Carcinoma
    Authors: YB Pang, J He, BY Cui, S Xu, XL Li, MY Wu, R Liang, Y Feng, X Guo, XH Zhang, XL Luo
    Stem Cells Int, 2019;2019(0):5680327.
    Species: Human
    Sample Types: Whole Cells
  6. Immunogenic neoantigens derived from gene fusions stimulate T cell responses
    Authors: W Yang, KW Lee, RM Srivastava, F Kuo, C Krishna, D Chowell, V Makarov, D Hoen, MG Dalin, L Wexler, R Ghossein, N Katabi, Z Nadeem, MA Cohen, SK Tian, N Robine, K Arora, H Geiger, P Agius, N Bouvier, K Huberman, K Vanness, JJ Havel, JS Sims, RM Samstein, R Mandal, J Tepe, I Ganly, AL Ho, N Riaz, RJ Wong, N Shukla, TA Chan, LGT Morris
    Nat. Med., 2019;0(0):.
    Species: Human
    Sample Types: Whole Cells
  7. Secretion of IFN-? Associated with Galectin-9 Production by Pleural Fluid Cells from a Patient with Extrapulmonary Tuberculosis
    Authors: J Zhao, B Shiratori, H Chagan-Yas, M Matsumoto, T Niki, M Tanaka, Y Takahashi, O Usami, Y Ashino, T Hattori
    Int J Mol Sci, 2017;18(7):.
    Species: Human
    Sample Types: Whole Cells
  8. 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 Types: Whole Cells
  9. Antigen-triggered interferon-gamma and interleukin-10 pattern in cured mucosal leishmaniasis patients is shaped during the active phase of disease.
    Authors: Nogueira R, Gomes-Silva A, Bittar R, Silva Mendonca D, Amato V, da Silva Mattos M, Oliveira-Neto M, Coutinho S, Da-Cruz A
    Clin Exp Immunol, 2014;177(3):679-86.
    Species: Human
    Sample Types: Whole Cells
  10. 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 Types: Whole Cells
  11. 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 Types: Whole Cells
  12. 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 Types: Whole Cells
  13. 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 Types: Whole Cells
  14. 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 Types: Whole Cells
  15. 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: Whole Cells
  16. 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 Types: Whole Cells
  17. 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 Types: Whole Cells
  18. 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 Types: Whole Cells
  19. 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 Types: Whole Cells
  20. 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 Types: Whole Cells
  21. 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 Types: Whole Cells
  22. Multiplex Genome Editing to Generate Universal CAR T Cells Resistant to PD1 Inhibition.
    Authors: Ren J, Liu X, Fang C, Jiang S, June C, Zhao Y
    Clin Cancer Res, 0;23(9):2255-2266.
    Species: Human
    Sample Types: Whole Cells

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Reviews for Human IFN-gamma ELISpot Kit

Average Rating: 4.6 (Based on 10 Reviews)

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Human IFN-gamma ELISpot Kit
By Anonymous on 08/20/2021
Application: Sample Tested: Human T cells

Human IFN-gamma ELISpot Kit
By Anonymous on 03/26/2021
Application: Sample Tested: Human T cell

Human IFN-gamma ELISpot Kit
By Anonymous on 02/09/2021
Application: Sample Tested: Human CD8 cells

Human IFN-gamma ELISpot Kit
By Anonymous on 09/30/2020
Application: Sample Tested: Activated human CD8 T cells

Works really well as compared to kits from other companies. Easy to follow protocol.


Human IFN-gamma ELISpot Kit
By Anonymous on 07/15/2020
Application: Sample Tested: Peripheral blood mononuclear cells (PBMCs)

The kit is easy to use and produced excellent results. Couple things that need to be added are: Information regarding storage conditions of the developed kit (spots); In my experience, the kit once developed is pretty stable and can be stored @ 4C in the dark for at least few weeks. The positive control for the kit produces a lawn rather than spots, something to keep in mind.


Human IFN-gamma ELISpot Kit
By Anonymous on 07/10/2020
Application: Sample Tested: Peripheral blood T cells

Human IFN-gamma ELISpot Kit
By Anonymous on 07/10/2020
Application: Sample Tested: Human cells

Human IFN-gamma ELISpot Kit
By Anonymous on 01/25/2020
Application: Sample Tested: Serum and Plasma

Human IFN-gamma ELISpot Kit
By Anonymous on 05/14/2018
Application: Sample Tested: Cell Culture Samples

Human IFN-gamma ELISpot Kit
By Anonymous on 03/22/2018
Application: Sample Tested: Human PBMC