Novex® multiplex assays are performed in much the same way as ELISAs (Figure 5). The steps are similar, with the exception that antibody-specific capture beads are added to the wells of 96-well microtiter plates, instead of having capture antibodies attached to the wells. Samples are then placed into the microtiter plate wells. Novex® multiplex assay kits are provided with protein standards of known concentration, so that standard curves for the proteins being analyzed can be generated.
After incubation, the beads are washed and resuspended in a solution containing biotinylated detection antibody. Another incubation and wash step is performed, followed by the addition of streptavidin–R-phycoerythrin (R-PE). The beads are then washed and are ready to be analyzedon a Luminex® instrument.