Mouse IFN-gamma/IL-4 Dual-Color ELISpot Kit

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Product Details
Citations (9)
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Mouse IFN-gamma/IL-4 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/IL-4. 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/IL-4. 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/IL-4.

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/IL-4 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/IL-4
  • 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/IL-4 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/IL-4 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/IL-4 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.

9 Citations: Showing 1 - 9
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  1. Effector/memory CD4 T cells making either Th1 or Th2 cytokines commonly co-express T-bet and GATA-3
    Authors: A Das, V Ranganatha, D Umar, S Thukral, A George, S Rath, V Bal
    PLoS ONE, 2017-10-31;12(10):e0185932.
    Species: Mouse
    Sample Types: Whole Cells
  2. Host- and pathogen-derived adjuvant coatings on protein nanoparticle vaccines
    Authors: TZ Chang, I Diambou, JR Kim, B Wang, JA Champion
    Bioeng Transl Med, 2017-02-03;2(1):120-130.
    Species: Mouse
    Sample Types: Whole Cells
  3. Yeast Surface-Displayed H5N1 Avian Influenza Vaccines
    Authors: H Lei, S Jin, E Karlsson, S Schultz-Ch, K Ye
    J Immunol Res, 2016-12-19;2016(0):4131324.
    Species: Mouse
    Sample Types: Whole Cells
  4. Effective inhibition of melanoma tumorigenesis and growth via a new complex vaccine based on NY-ESO-1-alum-polysaccharide-HH2.
    Authors: Li M, Shi H, Mu Y, Luo Z, Zhang H, Wan Y, Zhang D, Lu L, Men K, Tian Y, Wu X, Liu X, Pan Y, Fan Y, Yu C, Zhou B, Xiang R, Chen X, Yang L
    Mol Cancer, 2014-07-28;13(0):179.
    Species: Mouse
    Sample Types: Whole Cells
  5. Antitumor effects of L-BLP25 Antigen-Specific tumor immunotherapy in a novel human MUC1 transgenic lung cancer mouse model.
    Authors: Wurz G, Gutierrez A, Greenberg B, Vang D, Griffey S, Kao C, Wolf M, Degregorio M
    J Transl Med, 2013-03-13;11(0):64.
    Species: Mouse
    Sample Types: Whole Cells
  6. Plasmid DNA containing multiple CpG motifs triggers a strong immune response to hepatitis B surface antigen when combined with incomplete Freund's adjuvant but not aluminum hydroxide.
    Authors: Luo Z, Shi H, Zhang H, Li M, Zhao Y, Zhang J, Guo F, Luo S, Sun P, Zhang D, Qian Z, Yang L
    Mol Med Rep, 2012-09-12;6(6):1309-14.
    Species: Mouse
    Sample Types: Whole Cells
  7. An epigenetic silencing pathway controlling T helper 2 cell lineage commitment.
    Authors: Allan RS, Zueva E, Cammas F, Schreiber HA, Masson V, Belz GT, Roche D, Maison C, Quivy JP, Almouzni G, Amigorena S
    Nature, 2012-07-12;487(7406):249-53.
    Species: Mouse
    Sample Types: Whole Cells
  8. L-BLP25 Vaccine plus Letrozole Induces a TH1 Immune Response and Has Additive Antitumor Activity in MUC1-Expressing Mammary Tumors in Mice.
    Authors: Mehta NR, Wurz GT, Burich RA, Greenberg BE, Griffey S, Gutierrez A, Bell KE, McCall JL, Wolf M, Degregorio M
    Clin. Cancer Res., 2012-03-20;18(10):2861-71.
    Species: Mouse
    Sample Types: Whole Cells
  9. HOCl oxidation-modified CT26 cell vaccine inhibits colon tumor growth in a mouse model.
    Authors: Zhou, Rui, Huang, Wen-Jun, Ma, Cong, Zhou, Yan, Yao, Yu-Qin, Wang, Yu-Xi, Gou, Lan-Tu, Yi, Chen, Yang, Jin-Lian
    Asian Pac J Cancer Prev, 2012-01-01;13(8):4037-43.
    Species: Mouse
    Sample Types: Whole Cells


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