This method does not involve a live/dead stain therefore NK cells and tumor targets must have good viability (>90%) at the start of the assay for results to be meaningful.
If a drug is being used to increase adherence e.g. Rituximab, consider including the same cell line without the drug to evaluate the contribution of the drug.
We have tested this assay using K562 as a readout of natural cytotoxicity and Raji with Rituximab (Genentech) as a readout of antibody dependent cellular cytotoxicity. Effector:target ratios will impact the frequency of adherence. A 1:2 ratio used here gives NK cells a high chance of encountering a tumor cell. The experimental readout is the proportion of NK cells in conjugates. Increase the proportion of tumor cells to maximize the likelihood of NK cells encountering a tumor cell (and therefore the limiting factor is its ability to adhere to a target cell).
Effector cells will be able to adhere to different target types at different rates. When working with a new target cell or effector, optimize the effector:target cell ratio and timing for maximum sensitivity to experimental perturbation.