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- Yunfei Zhang, J Brian Robertson, Qiguang Xie, and Carl Hirschie Johnson.
- Department of Biological Sciences, Vanderbilt University, Nashville, TN, 37235, USA.
- Methods Mol. Biol. 2016 Jan 1; 1461: 117-30.
Abstract"pHlash" is a novel bioluminescence-based pH sensor for measuring intracellular pH, which is developed based on Bioluminescence Resonance Energy Transfer (BRET). pHlash is a fusion protein between a mutant of Renilla luciferase (RLuc) and a Venus fluorophore. The spectral emission of purified pHlash protein exhibits pH dependence in vitro. When expressed in either yeast or mammalian cells, pHlash reports basal pH and cytosolic acidification. In this chapter, we describe an in vitro characterization of pHlash, and also in vivo assays including in yeast cells and in HeLa cells using pHlash as a cytoplasmic pH indicator.
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