RatioWorks™ PH165, NHS ester

RatioWorks™ PH165 NHS ester 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. The amine-reactive RatioWorks™ PH165 NHS ester is an excellent building block for developing novel ratiometric fluorescent probes of a variety of biological applications that require the precise pH determinations with the minimal effect of pH probe concentration change.

The pH dependent absorbance spectra of  RatioWorks™ PH165.

Protocol

SDS

COA

Catalog	Size	Price	Quantity
21214	1 mg	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	581.51
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

Documents

Protocol
Safety Data Sheet
Certificate of Analysis

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Page updated on November 1, 2025