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Cell Meter™ Fluorimetric Intracellular pH Assay Kit
Intracellular pH change are implicated in diverse physiological and pathological processes, including cell proliferation, apoptosis, fertilization, malignancy, multidrug resistance, ion transport, lysosomal storage disorders and Alzheimer's disease. The Cell Meter™ Fluorimetric Intracellular pH Assay Kit utilizes AAT Bioquest's proprietary fluorescent indicator for measuring the relative intracellular pH changes. It is a homogeneous, kinetic, live-cell fluorescent assay that utilizes either a standard procedure or acid-load procedure. The standard protocol is designed for measuring the therapeutic targets of interest with a decrease in intracellular pH upon treatment. The 'Acid-Load' procedure is designed to measure the increase of intracellular pH associated with changes in cellular metabolism due to GPCR activation or growth factor activity. With the 'Acid-Load' procedure ammonium chloride solution is added after the fluorescent pH dye is loaded into cells in a minimum volume. This 'acid-loading' step is followed by the addition of agonist in a relatively large volume (~4X) of buffer. The sudden volume change initiates an efflux of ammonia (NH3) from the cells causing a rapid decrease in intracellular pH, and thus a decrease in fluorescence signal. The effect of agonist on the subsequent recovery of intracellular pH is measured by the relative fluorescence signal increase.
Carbachol dose response in CHO-M1 cell. CHO-M1 cells were seeded overnight in 60,000 cells per 100 µL per well in a 96- well black wall/clear bottom costar plate. The growth medium was replaced with 50 µL/well of RatioWorks™ BCFL, AM dye-loading solution for 37°C for 1 hour, follow by 15 minutes incubation with 5  µL/well of 220 mM NH4Cl. Carbachol (200µL/well) was added by FlexStation (Molecular Devices) to achieve the final indicated concentrations. The fluorescent signal was generated using Ex/Em = 490/535 nm (Cutoff = 515 nm).
Carbachol dose response in CHO-M1 cell. CHO-M1 cells were seeded overnight in 60,000 cells per 100 µL per well in a 96- well black wall/clear bottom costar plate. The growth medium was replaced with 50 µL/well of RatioWorks™ BCFL, AM dye-loading solution for 37°C for 1 hour, follow by 15 minutes incubation with 5  µL/well of 220 mM NH4Cl. Carbachol (200µL/well) was added by FlexStation (Molecular Devices) to achieve the final indicated concentrations. The fluorescent signal was generated using Ex/Em = 490/535 nm (Cutoff = 515 nm).
protocol
Protocol
sds
SDS
CatalogSize
Price
Quantity
211801000 Tests
Price
 
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Instrument settings
Fluorescence microplate reader
Excitation490 nm
Emission535 nm
Cutoff515 nm
Recommended plateBlack well/Clear bottom
Other instrumentsArrayScan, FDSS, FLIPR, FlexStation, IN Cell Analyzer, NOVOStar, ViewLux
Documents
Protocol
Safety Data Sheet
Certificate of Analysis
Brochure: Physiological Probes & Assay Kits
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Page updated on October 31, 2025