ELISpot and FluoroSpot: Antigen Specific Response Assays
Evaluating cell-mediated immune responses
We support global preclinical research with FluoroSpot and ELISpot testing—measuring cytokine-secreting cells at the single-cell level
ELISpot assays
ELISpot assays provide robust measurement of cytokine-secreting cells at the single cell level. Like an ELISA, antibodies for the target cytokine are coated onto a 96-well plate. The cells being assessed are cultured in the same wells, stimulated to release cytokine, and then removed. Bound cytokines are visualized as with an ELISA, using chemiluminescence, with each spot corresponding to a single secreting cell.
FluoroSpot assays
FluoroSpot assays are similar to ELISpot assays but rely on detection using fluorescently labeled detection antibodies.
Fluorescent detection enables simultaneous measurement of cells secreting different cytokines through multiplexing.
Example ELISpot assay panels
| Stimuli | Cell Type Simulated | Details | Stimulates Secretion of Human |
|---|---|---|---|
| CEF peptide pool | CD8+ T cells | Ag-specific stimulation by T cell epitopes derived from CMV, EBV, and influenza | IFN-γ, IL-2, Granzyme B, Perforin |
| CEFT peptide pool | CD4+ T cells | Ag-specific stimulation by T cell epitopes derived from CMV, EBV, Tetanus Toxin and influenza | IFN-γ, IL-2, IL-5, IL-13 |
| Anti-CD3 Ab | T cells | Polyclonal T cell activation | IFN-γ, 2, 4, 5, 10, 13, 17A |
| PHA | T cells | Polyclonal T cell activation | IFN-γ, IL-2, 4, 5, 6, 10, 13, 17A, 21, GM-CSF, Granzyme B, Perforin |
| R848 + IL-2 | B cells | Polyclonal B cell activation | IgG, IgA, IgM |
| Anti-CD40 + IL-4 | B cells | Polyclonal B cell activation | IgE |
| LPS | Monocytes/DC | TLR4 agonist | IL,1β, 6, 8, 10, 12, 23, GM-CSF, MIP1β, TNF-α |
| R848 | Monocytes/DC | TLR/8 agonist | IFN-α, IL-1β, 6, 23, GM-CSF, TNF-α |
Choosing the right immune monitoring technology
Understand use cases for cytokine profiling, cell-based immunophenotyping, gene expression, sequencing, circulating tumor markers, and immunohistochemistry and multiplex immunofluorescence.
Additional lab services
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Cytokine Profiling & Proteomics
ExploreCustomized assays to detect cytokines or proteins of interest utilizing ELISA, Luminex, and MSD.
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ExploreCustomized assays to detect cytokines or proteins of interest utilizing ELISA, Luminex, and MSD.
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