pHrodo™ AM Variety Pack

Measure and quantify intracellular pH with the pHrodo AM Variety Pack, which includes pHrodo Red and Green AM indicator dyes, fluorogenic probes that traverse the cell membrane and remain within the intracellular space upon cleavage by nonspecific esterases.

The pHrodo AM Variety Pack contains one vial each of the pHrodo Red AM and pHrodo Green AM indicator dyes, with each vial providing enough reagent volume to test one 96-well plate. pHrodo Red AM and pHrodo Green AM intracellular pH indicators are fluorogenic probes used to measure intracellular pH in live cells. They are weakly fluorescent at neutral pH, but dramatically increase in brightness as the pH decreases. These reagents can quantify cellular cytosolic pH in the range of 4–9 with a pKa of ∼6.5 and have excitation/emission spectra of 560/585 nm (pHrodo Red) and 509/533 nm (pHrodo Green).

pHrodo Red AM and pHrodo Green AM dyes are photostable and appropriate for use in multiplexing experiments; pHrodo Red easily multiplexes with a wide variety of blue, green, and far-red dyes and reporters such as GFP, Fuo-4, calcein, NucBlue, CellEvent Caspase 3/7 green, Mitosox Green, and Mitotracker Deep Red, among many others. pHrodo Green dye easily multiplexes with a wide variety of blue, red, and far-red dyes reporters such as Mitosox Red, CellEvent Caspase 3/7 Red, NucBlue, RFPs, and Mitotracker Deep Red, among many others. 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). These reagents are also compatible with widefield or confocal fluorescence microscopes, flow cytometers, fluorescent plate readers, and high content instruments.