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RatioWorks™ PH165

The pH dependent absorbance spectra of  RatioWorks™ PH165.
The pH dependent absorbance spectra of  RatioWorks™ PH165.
The pH dependent absorbance spectra of  RatioWorks™ PH165.
The pH dependent Emission spectra of RatioWorks™ PH165.
The pH dependent Excitation spectra of  RatioWorks™ PH165.
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Additional ordering information
Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
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ShippingStandard overnight for United States, inquire for international
Physical properties
Molecular weight445.56
SolventDMSO
Spectral properties
Excitation (nm)463
Emission (nm)641
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
UNSPSC12171501

OverviewpdfSDSpdfProtocol


Molecular weight
445.56
Excitation (nm)
463
Emission (nm)
641
RatioWorks™ PH165 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-impermeable, and compatible with various platforms such as fluorescence microscopy and microplate reader.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of RatioWorks™ PH165 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 mM224.437 µL1.122 mL2.244 mL11.222 mL22.444 mL
5 mM44.887 µL224.437 µL448.873 µL2.244 mL4.489 mL
10 mM22.444 µL112.218 µL224.437 µL1.122 mL2.244 mL

Molarity calculator

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Spectrum


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spectrum

Spectral properties

Excitation (nm)463
Emission (nm)641

Product Family


NameExcitation (nm)Emission (nm)
RatioWorks™ PDMPO333531

Images


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