New Application Report explains how to use fluorescence-activated cell sorting (FACS) to rescue low expression cells for efficient APC assays
Kv1.3 ion channels regulate T lymphocyte activation, controlling membrane potential and calcium signaling in immune responses. Dysregulation is linked to autoimmune diseases like multiple sclerosis, psoriasis, and rheumatoid arthritis. Kv1.3 channels also play roles in cancer cell proliferation and survival. Hence, Kv1.3 channels are considered promising targets for therapies in both autoimmune conditions and cancer.
Efficient ion channel assays require consistent, robust, functional membrane expression. Working with a low expressing Kv1.3 cell line, we used the Tyto cell sorter (Miltenyi Biotec) to sort cells based on their Kv1.3 expression to drastically improve success rates on QPatch assays.
Summarizing the application report:
- Low Kv1.3 expression in a CHO cell line led to a 20% success rate in QPatch Automated Patch Clamp (APC) assays.
- To address this, fluorescence-activated cell sorting (FACS) was used to enrich the cell population expressing the Kv1.3 ion channel.
- By staining cells with PE-labeled antibodies against the Kv1.3 channel and sorting based on fluorescence intensity, the APC success rate improved to 80%.
- This process did not affect cell viability, seal formation, or channel activity.
- Overall, FACS significantly enhances APC assay outcomes in low-expressing cell lines.