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CAS:117464-70-7
Appearance:White to off-white solid
Storage:Powder : -20℃, 1 years;In solvent(mother liquid): -20℃, 1 month; -80℃, 6 months
Purity:≥90%
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BCECF, AM is a fluorescent dye that can penetrate cell membranes to detect intracellular pH. BCECF, AM does not fluoresce, and can be clipped into BCECF by intracellular esterase after entering the cell, thus being retained in the cell. BCECF can be excited to form green fluorescence at the right pH value. The maximum excitation wavelength and emission wavelength are different due to the difference in pH, the maximum excitation wavelength is about 503nm, the maximum emission wavelength is about 520nm, and the recommended excitation wavelength is 488nm and the emission wavelength is 535nm.
BCECF and AM are not only widely used in the study of mammalian cells, but also reported to be used in the detection of intracellular pH levels in animal tissues, plant cells, bacteria and yeast. BCECF and AM are widely used in the process of cytotoxicity, apoptosis, cell adhesion, drug resistance, cell chemotaxis and other processes with intracellular pH changes.
When used for intracellular pH detection, the concentration of BCECF and AM commonly used is 1-10 μM.
Protocol (for reference only)
(1) Reagents
1 mM BCECF, AM/DMSO
HEPES buffer (20 mM HEPES, 153 mM NaCl, 5 mM KCl, 5 mM glucose, pH 7.4)
(2) Protocol
①. Cell suspension was prepared with HEPES and cell concentration was 4×107 cells /mL.
②. 1mM BCECF, AM/DMSO solution was added into the cell suspension (1/300 volume of the cell suspension), and the final concentration of BCECF and AM was 3μM.
③. Incubate at 37 oC for 30 min.
(4) Cells were washed with HEPES buffer 3 times to produce 3×106 cells /mL cell suspension.
(5) The fluorescence intensity of cells is detected by fluorescence microscopy or laser copolymerization microscopy with image analysis system.
* The conditions for labeling vary depending on the cell type, so please determine the best conditions before each experiment. The above methods are for reference only.
Note:
1) BCECF, AM may be harmful to human body, please pay attention to proper protection.
2) Fluorescent dyes have quenching problems, please pay attention to avoid light as much as possible to slow down fluorescence quenching.
