RatioWorks™ PH165, NHS ester

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Shipping	Standard overnight for United States, inquire for international

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

Overview SDS Protocol

Molecular weight

581.51

Excitation (nm)

463

Emission (nm)

641

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.

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of RatioWorks™ PH165, NHS ester to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	171.966 µL	859.83 µL	1.72 mL	8.598 mL	17.197 mL
5 mM	34.393 µL	171.966 µL	343.932 µL	1.72 mL	3.439 mL
10 mM	17.197 µL	85.983 µL	171.966 µL	859.83 µL	1.72 mL

Molarity calculator

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Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
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Spectrum

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Spectral properties

Excitation (nm)	463
Emission (nm)	641

Images

Figure 1. The pH dependent absorbance spectra of RatioWorks™ PH165.

Figure 2. The pH dependent Emission spectra of RatioWorks™ PH165.

Figure 3. The pH dependent Excitation spectra of RatioWorks™ PH165.

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

References

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