BCECF-AM Chemical Properties

Boiling point:

692.41°C (rough estimate)

Density 

1.2837 (rough estimate)

refractive index 

1.6500 (estimate)

Flash point:

85 °C

storage temp. 

-20°C

solubility 

acetonitrile: 25mg/mL
DMF: soluble
DMSO: soluble

form 

White to off-white solid

color 

Orange to red

Appearance

white powder

CAS DataBase Reference

117464-70-7(CAS DataBase Reference)

Safety Information

Hazard Codes 

Xi

Risk Statements 

36/37/38

Safety Statements 

22-26-36

WGK Germany 

3

MSDS

• Language:English Provider:ACROS

BCECF-AM Usage And Synthesis

Description

BCECF AM ester is a widely-used fluorescent indicator for estimating intracellular pH. This membrane-permeant compound is modified by intracellular esterases to produce BCECF, a polar fluorescein derivative that is retained by cells. Intracellular BCECF is stable and has an efflux half-life of greater than two hours. Intracellular pH measurements are made by ratio imaging, which involves determining the pH-dependent ratio of emission intensity (detected at 535 nm) when the dye is excited at 490 nm vs. the emission intensity when excited at 440 nm. This approach is amenable to either spectrofluorometry or flow cytometry. BCECF AM can also be used to investigate intracellular changes in other ions, including potassium.

Chemical Properties

pale yellow to yellow powder

Uses

Cell-permeable ester of BCECF that, on hydrolysis by cytosolic esterases, yields the intracellularly trapped pH-indicator BCECF. Allows simultaneous recording of cell volume changes and intracellular pH in single osteosarcoma cells.

General Description

Material is an isomeric mixture of lactones, acetates, and AM esters

Biological Activity

Non-charged, non-fluorescent pH indicator. Readily and rapidly diffuses through cell membranes.

References

[1] T J RINK  T P  R Y Tsien. Cytoplasmic pH and free Mg2+ in lymphocytes.[J]. Journal of Cell Biology, 1982, 95 1: 189-196. DOI: 10.1083/jcb.95.1.189
[2] PINAR OZKAN  Raj M. A rapid method for measuring intracellular pH using BCECF-AM[J]. Biochimica et biophysica acta. General subjects, 2002, 1572 1: Pages 143-148. DOI: 10.1016/s0304-4165(02)00303-3
[3] CAROLINE DIVE. Polar fluorescein derivatives as improved substrate probes for flow cytoenzymological assay of cellular esterases[J]. Molecular and Cellular Probes, 1988, 2 2: Pages 131-145. DOI: 10.1016/0890-8508(88)90035-7
[4] NATHAN O’CONNOR  Randi B S. Ratio imaging: practical considerations for measuring intracellular Ca2+ and pH in living cells.[J]. Methods in cell biology, 2013, 114: 387-406. DOI: 10.1016/b978-0-12-407761-4.00016-6
[5] SUE CHOW  Ian T  David Hedley. Flow cytometric calibration of intracellular pH measurements in viable cells using mixtures of weak acids and bases[J]. Cytometry Part B: Clinical Cytometry, 1996, 24 4: 360-367. DOI: 10.1002/(sici)1097-0320(19960801)24:4<360::aid-cyto7>3.0.co;2-j
[6] LáSZLó BALKAY. Flow cytometric determination of intracellular free potassium concentration[J]. Cytometry Part B: Clinical Cytometry, 1998, 28 1: 42-49. DOI: 10.1002/(sici)1097-0320(19970501)28:1<42::aid-cyto5>3.0.co;2-j

Abs/Em Maxima

440; 490/530 nm

Excitation / Emission Maximum

440-490 nm/535 nm

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