BCECF is the most widely used intracellular pH probe. BCECF is highly water-soluble because it has 4 to 5 negative charges at neutral pH value; it becomes difficult to pass through the cell membrane after loading. Its pKa value, 6.97, is higher than that of carboxyfluorescein. BCECF has an isosbestic point at 439 nm in the excitation spectra, so it can be used in ratiometry, similar to Fura 2.
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