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BCECF AM

BCECF AM (Ex-Max 503 nm/Em-Max 524 nm)

BCECF-AM is a cell-permeant pH indicator in its acetoxymethylester (AM) form with a pKa (6.97). This form of the pH indicator passively crosses the intact, live cell membrane where the AM ester moeity is cleaved by instracellular esterases, retaining the probe inside. At low pH, the reagent is weakly fluorescent but increases in intensity with increasing pH. This probe can be used ratiometrically as the excitation spectrum shifts between 439 nm and 505 nm, pH depending. This probe has, therefor, a primary excitation peak at 505 nm and a secondary peak at 439 nm with an emission peak that remains constant at 524 nm.

Excitation & emission spectra

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Key facts

Excitation maximum
503 nm
Emission maximum
524 nm
Optimal laser line
488
Optimal filter
515/30
Molecular weight
820.71

BCECF AM has an excitation maximum at 503 nm and an emission maximum at 524 nm, a Stokes shift of 21 nm. It is optimally excited by the 488 laser line.

Frequently asked questions

What is BCECF AM?
BCECF-AM is a cell-permeant pH indicator in its acetoxymethylester (AM) form with a pKa (6.97). This form of the pH indicator passively crosses the intact, live cell membrane where the AM ester moeity is cleaved by instracellular esterases, retaining the probe inside. At low pH, the reagent is weakly fluorescent but increases in intensity with increasing pH. This probe can be used ratiometrically as the excitation spectrum shifts between 439 nm and 505 nm, pH depending. This probe has, therefor, a primary excitation peak at 505 nm and a secondary peak at 439 nm with an emission peak that remains constant at 524 nm.
What laser excites BCECF AM?
BCECF AM is optimally excited by the 488 laser line. Its excitation maximum is 503 nm.
What filter should I use to detect BCECF AM?
A 515/30 filter captures the peak of the BCECF AM emission near 524 nm with minimal spillover.