Description
Aviva's Murine IFN gamma ELISpot Kit is based on standard sandwich ELISA methodology. Capture antibodies highly specific for the analyte of interest are coated to the wells of a PVDF bottomed 96 well microtitre plate either during kit manufacture or in the laboratory. The plate is then blocked to minimise any non-antibody dependent unspecific binding and finally washed before adding the cells to be investigated. Cell suspension and stimulant are added to the coated and blocked microtitre plate and the plate incubated allowing the specific antibodies to bind any analytes produced. Biotinylated detection antibodies are then added which bind to the previously captured analyte. Enzyme conjugated Streptavidin is added binding to the detection antibodies. Any excess unbound analyte and antibodies are removed by careful washing. Colour substrate is then applied to the wells resulting in coloured spots which can be quantified using appropriate analysis software or manually using microscopes