A Protocol for Measurement of Intracellular pH   

Edited by
Lin Fang
Reviewed by
Anonymous reviewer
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Original research article

A brief version of this protocol appeared in:
Molecular Cell
Jan 2013


Intracellular pH (pHi) is an important physiological determinant of enzyme activity and cellular function (Kurkdjian and Guern, 1989). All proteins depend on a tightly regulated pH to maintain their structure and function. Protonation–deprotonation events can dictate the charge of biological surfaces and are integral steps in many metabolic reactions (Casey et al., 2010). Moreover, the proton gradient across the mitochondrial membrane is used to generate cellular energy and support other mitochondrial processes. As a result, cells have developed multiple mechanisms to maintain a narrow range of pHi in response to extra- and intracellular fluctuations in pH (Orij et al., 2012). Here, we describe a protocol for pHi measurement in live cells that uses fluorescent microscopy and the pH sensitive dye 2’,7’-Bis-(2-Carboxyethyl)-5-(and-6-)-Carboxyfluorescein Acetoxymethyl Ester (BCECF-AM). This method was recently used to determine the effects of intracellular pH changes on global histone acetylation levels (McBrian et al., 2013).

Copyright: © 2014 The Authors; exclusive licensee Bio-protocol LLC.
How to cite: Behbahan, I. S., McBrian, M. A. and Kurdistani, S. K. (2014). A Protocol for Measurement of Intracellular pH. Bio-protocol 4(2): e1027. DOI: 10.21769/BioProtoc.1027.

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amgad abdelrahman
veterinary research institute
i hope to know the protocol of measuring mineral of epithelial cell (hydrogen. phosphorus.carbon .sodium...etc)
1/13/2015 11:37:50 PM Reply
Bio-protocol team Bio-protocol team

Thank you for your suggestions. Our editorial committee will invite authors to contribute the requested protocols to our site in the near future.

1/20/2015 3:49:17 PM