Abstract
Calcium influx through store-operated Ca2+ entry (SOCE), mediated by STIM-operated Orai channels, is crucial for many cellular functions. To dissect the molecular mechanisms underlying the process of STIM-Orai activation and identify regulators that modify this process, ratiometric imaging of SOCE responses in HEK cells overexpressing STIM and Orai is a routinely used method. Here we describe one commonly used procedure of monitoring SOCE activity with a ratiometric membrane-permeable dye fura-2-AM. Other ratiometric indicators suitable for SOCE measurements are also discussed.
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Acknowledgments
This work was supported by the National Natural Science foundation of China (NSFC-31471279 and NSFC-31671492 to Y.W.), the Recruitment Program for Young Professionals of China (to Y.W.), grants from NIH (R21GM126532 and R01GM112003 to Y.Z.), CPRIT (RP170660, to Y.Z.), the Welch Foundation (BE-1913 to Y.Z.) and the American Cancer Society (RSG-16–215-01-TBE to Y.Z.).
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Zhang, S., He, L., Zhou, Y., Wang, Y. (2018). Fluorescence-Based Ratiometric Measurement of CRAC Channel Activity in STIM-Orai-Overexpressing HEK-293 Cells. In: Penna, A., Constantin, B. (eds) The CRAC Channel. Methods in Molecular Biology, vol 1843. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8704-7_2
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DOI: https://doi.org/10.1007/978-1-4939-8704-7_2
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