Monitoring Intracellular pH Change with a Genetically Encoded and Ratiometric Luminescence Sensor in Yeast and Mammalian Cells.
Methods Mol Biol
; 1461: 117-30, 2016.
Article
em En
| MEDLINE
| ID: mdl-27424899
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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Base de dados:
MEDLINE
Assunto principal:
Técnicas Biossensoriais
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Espaço Intracelular
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Concentração de Íons de Hidrogênio
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Medições Luminescentes
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Proteínas Luminescentes
Limite:
Animals
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Humans
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article