What are possible reasons for inconsistent Fluo-4 AM loading?

Cell types vary in their requirements. Incorrectly-prepared or unhealthy cell populations will not behave consistently. You may need to adjust your preparation or protocol to reflect these variations. Using a kit with recommended protocol may help with reproducibility.

Another common cause for inconsistent loading is that the cell medium may be contaminated or unsuitable for sample. Be sure to use serum-free medium to avoid unexpected esterase activity. For cell medium recipes and formulations, see our interactive tool in Additional Resources below.