The pHrodo® AM Variety Pack contains one vial each of pHrodo® Red AM and pHrodo® Green AM, enough to test one 96 well plate each. The pHrodo® Red AM and pHrodo® Green AM intracellular pH indicators are novel fluorogenic probes used to measure intracellular pH in live cells. They are weakly fluorescent at neutral pH but increasingly fluorescent as the pH drops. These reagent can quantify cellular cytosolic pH in the range of 9-4 with a pKa of ~6.5 and have excitation/emission of 560/585 nm (Red) and 509/533 nm (Green). pHrodo® Red AM and pHrodo® Green AM are photostable and appropriate for use in multiplexing experiments.
The pHrodo® Red and pHrodo® Green dyes have been modified with AM ester groups, which results in an uncharged molecule that can permeate cell membranes. Once inside the cell, the lipophilic blocking groups are cleaved by nonspecific esterases, resulting in a compound that is retained within the intracellular space. Fluorescence intensity of the probe is then an indicator of intracellular pH. Subsequent use of the Intracellular pH Calibration Buffer Kit (Cat. No. P35379) allows this intracellular pH to be quantified.
pHrodo® Red AM and pHrodo® Green AM intracellular pH indicators are compatible with various platforms, such as traditional fluorescence microscopy, high content screening (HCS), flow cytometry, and microplate-based fluorimetry or high throughput screening (HTS). This reagent is also compatible with various benchtop instruments such as the Evos®, FLoid™, Tali™, and Attune™ instruments.
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For Research Use Only. Not for use in diagnostic procedures.