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PIP2 Assays

PIP2 Assays

PIP2, also known as Phosphatidylinositol 4,5-bisphosphate or PtdIns(4,5)P2, is a substrate for several signaling molecules. It is a precursor for the DAG/IP3 pathway, but also plays a key role in phospholipid signaling and is involved in the regulation of ion channels and transporters.

Live cell PIP2 assays use a dimerization-dependent fluorescent protein sensor under the control of a CMV promoter. When PIP2 is hydrolyzed, fluorescence decreases. These fluorescent sensors can be combined with different colored sensors (red GECO calcium or DAG sensors, green cAMP or DAG sensors) to measure multiple signals simultaneously. PIP2 Assays are genetically-encoded and packaged in BacMam for easy transduction into a wide variety of cell types, including primary and iPSC-derived cell lines. Simply add to your cells and measure robust signals the very next day.

pip2fig1

Green fluorescent PIP2 sensor in HEK 293 cells, co-transduced with human muscarinic acetylcholine control receptor M1, and a red nuclear label. Fluorescence was captured from cells before the addition of 50 μM carbachol, and sampled at regular intervals. The maximal response is reached at ~60 seconds after the addition of the drug, and the response begins its return to baseline ~100 seconds after drug is added.

pip2fig2

Real-time live cell multiplex detection: A) fluorescent imaging with PIP2 and Upward DAG Assays.  B) simultaneous measurement of PIP2 and DAG (top: Upward DAG Assay; bottom: Downward DAG Assay).

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Applications: live cell assay, PIP2 hydrolysis measurements
Applications: live cell assay, PIP2 hydrolysis measurements