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Functional Studies - Intracellular pH Assay Kit (AB228552)
Carbachol dose response in CHO-M1 cells.
CHO-M1cells 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 dyeloading 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 (cut off at 525 nm).
Product details
The Intracellular pH Assay Kit (ab228552) utilizes a proprietary fluorescent indicator for measuring relative intracellular pH changes. It is an 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.
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.
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Publications (5)
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Molecular and cellular biochemistry : PubMed40588665
2025
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American journal of cancer research 13:4721-4733 PubMed37970357
2023
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Cells 11: PubMed36359862
2022
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Biochemistry and biophysics reports 27:101054 PubMed34189282
2021
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Oncotarget 12:859-872 PubMed33889306
2021
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Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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