Zinc (Zn) plays roles in structure, catalysis, and signaling. The majority of cellular Zn is bound by proteins, but a fraction of total Zn exists in a labile form. Here, we present a protocol for measuring labile cytosolic Zn using an in situ calibration of a genetically encoded Förster resonance energy transfer (FRET) sensor. We describe steps for producing buffered Zn solutions for performing an imaging-based calibration and analyzing the imaging data generated to determine labile Zn concentration in single cells. For complete details on the use and execution of this protocol, please refer to Rakshit and Holtzen et al..
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| http://dx.doi.org/10.1016/j.xpro.2024.103130 | DOI Listing |
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