RatioWorks™ PH165, AM

RatioWorks™ PH165 AM is a novel fluorescent probe that can be used to measure intracellular pH in live cells. It shows pH-dependent fluorescence spectra. Its dual excitation and dual emission make it a unique ratiometric pH probe for analyzing live cells. At low pH, RatioWorks™ PH165 is weakly fluorescent in far red spectrum region and highly fluorescent in red spectrum region. As pH increases, it shows higher fluorescence in far red spectrum and lower fluorescence in red spectrum. This probe can quantify cytosolic pH in the range from 4 to 9 with dual excitation/emission at 497/594 nm and 578/654 nm respectively. The pKa of PH165 is ~7.1. The red fluorescence spectrum of RatioWorks™ PH165 makes it suitable for multiplexing tests with green fluorophores such as GFP. RatioWorks™ PH165 AM is cell-permeable, and compatible with various platforms such as fluorescence microscopy and microplate reader.

Protocol

SDS

COA

Catalog	Size	Price	Quantity
21212	10x50 ug	Price

Citations

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

Physical properties

Molecular weight	583.53
Solvent	DMSO

Spectral properties

Excitation (nm)	463
Emission (nm)	641

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12171501

Instrument settings

Fluorescence microscope
Excitation	Cy3-Cy5 filter set
Emission	Cy3-Cy5 filter set
Recommended plate	Black wall/clear bottom
Instrument specification(s)	Cy3-Cy5 filter set

Fluorescence microplate reader
Excitation	497-578 nm
Emission	594-654 nm
Cutoff	570-630 nm
Recommended plate	Black wall/clear bottom
Instrument specification(s)	Bottom read mode

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Page updated on October 12, 2025