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Standard curve created using pHrodo™ Green AM with Intracellular pH Calibration Buffer Kit
U2OS cells were incubated with 10 mM pHrodo™ Green AM for 30 minutes at room temperature (with PowerLoad). The Intracellular pH Calibration Buffer Kit (Cat. No. P35379) was used to clamp the intracellular pH with extracellular buffer at pH 4.5, 5.5, 6.5 and 7.5. Intracellular pH vs. relative fluorescence units was plotted using a high content screening (HCS) instrument. An average of 3 data points was plotted and a linear trendline was fitted to obtain the pH standard curve. +/- error bars represent the standard deviation.
