Mouse IFN-gamma/Granzyme B Dual-Color ELISpot Kit

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R&D Systems ELISpot and FluoroSpot Kits
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Citations (5)
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Mouse IFN-gamma/Granzyme B Dual-Color ELISpot Kit Summary

Assay Type
Quantitative Sandwich ELISA
96-well PVDF-backed microplate
Assay Length
3 hours 35 mins to 4 hours 50 mins*
Sample Type
Whole Cells
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.

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

Appropriately stimulated cells are pipetted directly into the wells of a PVDF-backed microplate. The PVDF membrane has been pre-coated with capture antibodies specific for IFN-gamma/Granzyme B. The microplate is then placed in a humidified 37 °C CO2 incubator for the desired period of time. During this incubation period, the immobilized capture antibodies in the immediate vicinity of the secreting cells bind secreted IFN-gamma/Granzyme B.

At the desired time point, one analyte is detected using a biotinylated detection antibody and alkaline-phosphatase (AP)-conjugated to streptavidin. The second analyte is detected using a horseradish peroxidase (HRP)-conjugated detection antibody. Following incubation with the chromogens BCIP/NBT (substrate for AP, blue-black precipitate) and AEC (substrate for HRP, red precipitate), spots form to indicate the presence of IFN-gamma/Granzyme B secreting cells. Each spot represents an individual secreting cell. The spots can be counted with an ELISpot reader system or using a stereomicroscope.


  • Detect and quantitate individual cells secreting IFN-gamma/Granzyme B
  • 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



  • Mouse IFN-gamma/Granzyme B Microplate
  • Biotin-conjugated Detection Antibody
  • HRP-conjugated Detection Antibody
  • Streptavidin-conjugated to Alkaline Phosphatase
  • Dilution Buffers
  • Wash Buffer Concentrate
  • BCIP/NBT Chromogen
  • AEC Chromogen
  • Mouse IFN-gamma/Granzyme B Positive Controls


  • 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

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Preparation and Storage

The product is shipped with polar packs. 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.

Citations for Mouse IFN-gamma/Granzyme B Dual-Color 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.

5 Citations: Showing 1 - 5
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  1. Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice
    Authors: G Wang, X Wu, G Zhu, S Han, J Zhang
    Aging (Albany NY), 2020-03-22;12(6):5318-5335.
    Species: Mouse
    Sample Types: Whole Cells
  2. Autologous iPSC-Based Vaccines Elicit Anti-tumor Responses In�Vivo
    Authors: NG Kooreman, Y Kim, PE de Almeida, V Termglinch, S Diecke, NY Shao, TT Wei, H Yi, D Dey, R Nelakanti, TP Brouwer, DT Paik, I Sagiv-Barf, A Han, PHA Quax, JF Hamming, R Levy, MM Davis, JC Wu
    Cell Stem Cell, 2018-02-15;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
  3. Rad52 deficiency decreases development of lung squamous cell carcinomas by enhancing immuno-surveillance
    Authors: R Lieberman, J Pan, Q Zhang, M You
    Oncotarget, 2017-05-23;8(21):34032-34044.
    Species: Mouse
    Sample Types: Whole Cells
  4. Smac mimetics synergize with immune checkpoint inhibitors to promote tumour immunity against glioblastoma
    Authors: ST Beug, CE Beauregard, C Healy, T Sanda, M St-Jean, J Chabot, DE Walker, A Mohan, N Earl, X Lun, DL Senger, SM Robbins, P Staeheli, PA Forsyth, T Alain, EC LaCasse, RG Korneluk
    Nat Commun, 2017-02-15;8(0):.
    Species: Mouse
    Sample Types: Whole Cells
  5. CTLA4 aptamer delivers STAT3 siRNA to tumor-associated and malignant T cells.
    Authors: Herrmann A, Priceman S, Swiderski P, Kujawski M, Xin H, Cherryholmes G, Zhang W, Zhang C, Lahtz C, Kowolik C, Forman S, Kortylewski M, Yu H
    J Clin Invest, 2014-06-02;124(7):2977-87.
    Species: Mouse
    Sample Types: Whole Cells


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