Rescuing APC assay on low expressing cell line using fluorescence-based cell sorting

Live CHO-KV1.3 cells were stained using a PE-labelled secondary antibody and a primary antibody directed against the extracellular epitope of the KV1.3 ion channel. The stained cells were successively sorted based on the fluorescence intensity, and hence on the channel expression, and tested on QPatchII.

Here we demonstrate that:

• Sorting CHO-KV1.3 cells based on channel expression prior to APC experiments can substantially increase the final success rate (SR) from an average of 20% to an average of 80%.
• The staining and sorting procedure do not affect the quality of CHO-KV1.3 cells for APC experiments, in terms of viability, seal formation, and channel activity.
• Cells can be recultured after sorting, however with a relapsing ion channel expression over time.