Alternate Names:BCECF/AM Also known as 3′,6′-Bis(acetyloxy)-5(or 6)-[[(acetyloxy)methoxy]carbonyl]-3-oxo-bis[(acetyloxy)methyl] Ester Spiro[isobenzofuran-1(3H),9′-[9H]xanthene]-2′,7′-dipropanoic Acid; SPIRO(ISOBENZOFURAN-1(3H),9′-(9H)XANTHENE)-2′,7′-DIPROPANOIC ACID
Application:BCECF/AM is a readily membrane-permeable fluorescent indicator for the measurement of cytoplasmic pH
Molecular Weight:Average = 688.59
Molecular Formula:mixture of C30H20O11, C35H28O15 and C40H36O19
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.
* Refer to Certificate of Analysis for lot specific data (including water content).
BCECF/AM, a membrane-permeable fluorescent indicator for the measurement of cytoplasmic pH with an Abs/Em = 503/520 nm. BCECF/AM readily and rapidly diffuses through cell membranes. BCECF/AM is an acetoxymethyl ester of BCECF, is a variable mixture of cell-permeable ester derivatives of BCECF that are hydrolyzed by cytosolic esterases to yield intracellularly trapped indicator BCECF. BCECF/AM is a selective vacuolar accumulation of the fluorescent pH-indicator used to monitor intracellular pH changes in mammalian fibroblasts, gastric cells, lymphocytes, myocytes, and distal convoluted tubules.
1. Weiner, I.D., et al. 1989. Am. J. Physiol. 256: F957-F964. PMID: 2719125 2. Miyata, H., et al. 1989. Biochem. Biophys. Res. Commun. 163: 500-505. PMID: 2775282 3. Hayashi, H., et al. 1992. Am. J. Physiol. 262: C628-C634. PMID: 1550207 4. Sasaki, S., et al. 1992. Biochim. Biophys. Acta. 1137: 45-51. PMID: 1327152 5. Futsaether, C.M., et al. 1993. Can. J. Microbiol. 39: 180-186. PMID: 8467419
Cell staining for detecting cytoplasmic pH.
• Suspend cells in HEPES buffer solution (sc-300789) to prepare 4x107 cells/ml.
• Add 1 mM BCECF-AM/DMSO solution to the cell suspension to prepare 3 uM BCECF-AM (1/300 vol of cell suspension) as the final concentration.
• Incubate the cell suspension at 37° C for 30 min.
• Wash the cells 3 times with HEPES buffer saline, and then prepare 3x106 cells/ml of the cell suspension.
•Determine the fluorescence intensity using a fluorescence microscope or a confocal laser microscope coupled with an image analyzer.
NOTE: Cell staining conditions differ by cell type, so it is necessary to optimize the conditions for each experiment.
Soluble in acetone, methanol, DMSO, and DMF. Insoluble in water.
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Various publications cite BCECF/AM as a membraneVarious publications cite BCECF/AM as a membrane-permeable fluorescent indicator for the measurement of cytoplasmic pH. -SCBT Publication Review
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