The B-cell FluoroSpot is as sensitive as ELISpot but offers the possibility to, in the same well, separately define and enumerate B cells secreting antibodies of different isotypes. This can be done for antigen-specific B-cells or with the purpose to determine the total number of B cells producing different immunoglobulin isotypes.
In antigen-specific FluoroSpot, the antigen is immobilized to the PVDF membrane. Antibodies from B cells secreting different antigen-specific Ig (e.g. IgG or IgA) are detected using fluorophore-labeled isotype-specific detection antibodies making spots appear either as red (e.g IgG) or green (e.g. IgA).
When determining the total number of isotype-specific antibodies (e.g IgG and IgA), secreted IgG and IgA are bound by specific capture antibodies immobilized to the PVDF membrane. Detection is performed in the same manner as in antigen-specific B-cell FluoroSpot.
Antigen-specific IgG/IgA FluoroSpot
Download our B-cell ELISpot/FluoroSpot Brochure as a PDF
Total IgG/IgA FluoroSpot