Determine the frequency of CD8α+ cells releasing human IFN-γ
A Human CD8α+/IFN-γ ELISpot Kit (Catalog # EL3094) is now available from R&D Systems. The ELISpot method has been adapted to determine the frequency of CD8α+ cells releasing human IFN-γ. In this assay format, the PVDF-backed microplates are coated with both anti-human CD8α+ and anti-human IFN-γ monoclonal antibodies. CD8α+ cells are selectively captured on the plate. The ELISpot plate with CD8α+-enriched cells is placed in a 37° C CO2 incubator. During the incubation, anti-human IFN-γ antibodies capture IFN-γ secreted primarily by CD8α+ cells. A biotinylated polyclonal antibody specific for human IFN-γ detects the captured IFN-γ. The BCIP/NBT detection system results in a blue-black colored precipitate forming at the sites of IFN-γ localization, with each individual spot representing an individual IFN-γ secreting cell.
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