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CytoFix™ BCECF, AM *Optimized for long term cellular pH tracking*

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Physical properties
Molecular weight638.63
Spectral properties
Extinction coefficient (cm -1 M -1)90000
Excitation (nm)504
Emission (nm)527
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure


Molecular weight
Extinction coefficient (cm -1 M -1)
Excitation (nm)
Emission (nm)
Intracellular pH plays an important modulating role in many cellular events, including cell growth, calcium regulation, enzymatic activity, receptor-mediated signal transduction, ion transport, endocytosis, chemotaxis, cell adhesion and other cellular processes. pH-sensitive fluorescent dyes have been widely applied to monitor changes in intracellular pH in recent years. Imaging techniques that use fluorescent pH indicators also allow researchers to investigate these processes with much greater spatial resolution and sampling density that can be achieved using other technologies such as microelectrode. Among them, 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF) is the most popular pH probe since it can be used to monitor cellular pH ratiometrically with pKa ~7.0. BCECF AM is the cell-permeable version of BCECF. However, BCECF rapidly leaks out of lives cell when BCECF is loaded into live cells through its AM ester due to the activities of drug efflux transporters. CytoFix™ BCECF is a modified BCECF derivative that contains a cell-retention group. The newly modified BCECF derivative is well retained in live cells for much longer time than the parent BCECF dye. CytoFix™ BCECF retains the same pH profile of BCECF with improved cell retention.


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of CytoFix™ BCECF, AM *Optimized for long term cellular pH tracking* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM156.585 µL782.926 µL1.566 mL7.829 mL15.659 mL
5 mM31.317 µL156.585 µL313.17 µL1.566 mL3.132 mL
10 mM15.659 µL78.293 µL156.585 µL782.926 µL1.566 mL

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Spectral properties

Extinction coefficient (cm -1 M -1)90000
Excitation (nm)504
Emission (nm)527


View all 3 citations: Citation Explorer
Intracellular pH Measurements in Glioblastoma Cells Using the pH-Sensitive Dye BCECF
Authors: Galifianakis, N. V., Placantonakis, D. G., Chesler, M.
Journal: Methods Mol Biol (2018): 103-109
Evaluation of BCECF fluorescence ratio imaging to properly measure gastric intramucosal pH variations in vivo
Authors: Rochon, P., Jourdain, M., Mangalaboyi, J., Fourrier, F., Soulie-Begu, S., Buys, B., Dehlin, G., Lesage, J. C., Chambrin, M. C., Mordon, S.
Journal: J Biomed Opt (2007): 064014
Calibration methods and avoidance of errors in measurement of intracellular pH (pHcyt) using the indicator bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) in human platelets
Authors: Valant, P. A., Haynes, D. H.
Journal: J Fluoresc (1992): 191-206


View all 1 references: Citation Explorer
Estimation of intramitochondrial pca and ph by fura-2 and 2,7 biscarboxyethyl-5(6)-carboxyfluorescein (bcecf) fluorescence
Authors: Davis MH, Altschuld RA, Jung DW, Brierley GP.
Journal: Biochem Biophys Res Commun (1987): 40