Figure 18. Stimulation assay workflow and plate setup.

A. Schedule of stimulation assay experiments, beginning with thawing all 5KC-TCR reporter cell lines and their corresponding K562-APC lines at day 0. Stimulation assays must be set up no earlier than 4 days and no later than 7 days after 5KC-TCR reporter cells are thawed. Stimulation assay plate should be read between 18 and 32 h after the addition of 5KC-TCR cells to ensure sufficient time for response to be mounted. B. Workflow of stimulation plate setup on Day 4. Note that if antigens are proteins, rather than short peptides, K562-APCs and antigens should be cultured overnight before adding 5KC-TCR reporter cells.