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Phospholipase C produces two second messengers - diacylglycerol (DAG), which remains in the membrane, and inositol triphosphate (IP 3 ), which triggers the release of calcium ions (Ca 2+ ) from intracellular stores. Genetically encoded sensors based on a single circularly permuted fluorescent protein (FP) are robust tools for studying intracellular Ca 2+  dynamics. We have developed a robust sensor for DAG based on a circularly permuted green FP that can be co-imaged with the red fluorescent Ca 2+  sensor R-GECO for simultaneous measurement of both second messengers.

Simultaneous Detection of Ca2+ and Diacylglycerol Signaling in Living Cells