The PAIA assay kits contain PAIAplates with dried capture beads and a reaction buffer containing the fluorescence marker. All PAIA assays follow the same general workflow:
First, the reaction buffer is dispensed into the wells of the PAIA plate. Afterwards the samples and the calibration standards are added. It is recommended to prepare at least ten calibration standards, covering the concentration range of the assay and ideally diluted in the same matrix as the samples. (Depending on to the expected protein concentration, between 2 and 10µL of sample are added)
The loaded PAIA plate is then placed on a shaker until the binding between capture beads, analyte and fluorescence marker is completed. Depending on the assay format, this takes between 15 and 40 minutes. Afterwards the PAIA plate is removed from the shaker and beads are allowed to settle for at least 10 minutes on the bench prior to measuring the plate.
Please take five minutes to look at the video tutorial!