BCECF/AM (CAS 117464-70-7)

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Application:	A membrane-permeable fluorescent indicator for the measurement of cytoplasmic pH
CAS Number:	117464-70-7
Purity:	≥90%
Molecular Weight:	Average = 688.58
Molecular Formula:	mixture of C₃₀H₂₀O₁₁, C₃₅H₂₈O₁₅ and C₄₀H₃₆O₁₉

Description

BCECF/AM, a membrane-permeable fluorescent indicator for the measurement of cytoplasmic pH with an Abs/Em = 503/520 nm. The compound 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.

Usage

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.

Technical Information

Appearance:	White to off-white
Physical State:	Solid
Solubility:	Soluble in acetone, methanol, DMSO, and DMF. Insoluble in water.
Storage:	Store at -20° C
Boiling Point:	637.07 °C at 760 mmHg (Predicted)
Density:	1.37 g/cm³ (Predicted)
Refractive Index:	n²⁰_D 1.69 (Predicted)
Fluoresence:	λex 505 nm, λem 520 nm in DMSO
IC50:	diacylglycerol kinase in isolated platelet membrane: IC₅₀ = 300 nM; Inhibition of diacylglycerol kinase in intact platelets: IC₅₀ = 120 nM; mitochondrial import inner membrane translocase subunit TIM23: IC₅₀ = 5440 nM (Saccharomyces cerevisiae S288c)

Safety and Reference Information

WGK Germany:

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.

References

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

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