BCECF-AM
Catalog No.: GC14603Purity:>95.00%Synonyms: BCECF Acetoxymethyl ester
Intracellular ratiometric pH indicator
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 117464-70-7
1mgIn Stock
$239.00
1
5mgIn Stock
$1,133.00
1
| Pack Size | Price | Availability | Quantity | Action |
|---|---|---|---|---|
| 1mg | $239.00 | In Stock | 1 | |
| 5mg | $1,133.00 | In Stock | 1 |
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Citations
1 publicationsPublications from researchers using this productDescription
BCECF AM ester is a widely-used fluorescent indicator for estimating intracellular pH.[1],[2] 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.[3] 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.[2],[4] This approach is amenable to either spectrofluorometry or flow cytometry.[2],[5] BCECF AM can also be used to investigate intracellular changes in other ions, including potassium.[6]
Reference:
[1]. Rink, T.J., Tsien, R.Y., and Pozzan, T. Cytoplasmic pH and free Mg2+ in lymphocytes. Journal of Cell Biology 95(1), 189-196 (1982).
[2]. Ozkan, P., and Mutharasan, R. A rapid method for measuring intracellular pH using BCECF-AM. Biochimica et Biophysica Acta 1572(1), 143-148 (2002).
[3]. Dive, C., Cox, H., Watson, J.V., et al. Polar fluorescein derivatives as improved substrate probes for flow cytoenzymological assay of cellular esterases. Mol.Cell Probe 2(2), 131-145 (1988).
[4]. O'Connor, N., and Silver, R.B. Ratio imaging: Practical considerations for measuring intracellular Ca2+ and pH in living cells. Methods Cell Biol. 81, 415-433 (2007).
[5]. Chow, S., Hedley, D., and Tannock, I. Flow cytometric calibration of intracellular pH measurements in viable cells using mixtures of weak acids and bases. Cytometry 24, 360-367 (1996).
[6]. Balkay, L., Márián, T., Emri, M., et al. Flow cytometric determination of intracellular free potassium concentration. Cytometry 28, 42-49 (1997).
Protocol
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Cell experiment: |
PASMCs are placed in a laminar flow cell chamber perfused with HBSS with pH adjusted to 7.4. pHi is measured in cells incubated with the membrane permeant (acetoxymethyl ester) form of the pH-sensitive fluorescent dye BCECF-AM for 60 min at 37°C under an atmosphere of 20% O2-5% CO2. Cells are then washed with HBSS for 15 min at 37°C to remove extracellular dye and allow complete de-esterification of cytosolic dye. Ratiometric measurement of BCECF fluorescence is performed on a workstation consisting of a Nikon TSE 100 Ellipse inverted microscope with epi-fluorescence attachments. The light beam from a xenon arc lamp is filtered by interference filters at 490 and 440 nm, and focused onto the PASMCS under examination via a 20× fluorescence objective. Light emitted from the cell at 530 nm is returned through the objective and detected by an imaging camera. An electronic shutter is used to minimize photobleaching of dye. Protocols are executed and data collected on-line with InCyte software. pHi is estimated from in situcalibration after each experiment. Cells are perfused with a solution containing (in mM): 105 KCl, 1 MgCl2, 1.5 CaCl2, 10 glucose, 20 HEPES-Tris and 0.01 nigericin to allow pHi to equilibrate to external pH. A two point calibration is created from fluorescence measured as pHi is adjusted with KOH from 6.5 to 7.5. Intracellular H+ ion concentration ([H+]i) is determined from pHi using the formula: pHi = −log ([H+]i). |
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References: [1]. Clark Undem, et al. Endothelin-1 Augments Na+/H+ Exchange Activity in Murine Pulmonary Arterial Smooth Muscle Cells via Rho Kinase. PLoS One. 2012; 7(9): e46303. |
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Product Documents
Purity:>95.00%Appearance:An oil
Chemical Properties
CAS No.
117464-70-7
Synonyms
BCECF Acetoxymethyl ester
Chemical Name
3',6'-bis[(acetyloxy)methoxy]-5(or 6)-[[(acetyloxy)methoxy]carbonyl]-3-oxo-spiro[isobenzofuran-1(3H),9'-[9H]xanthene]-2',7'-dipropanoic acid, 2',7'-bis[(acetyloxy)methyl] ester
SMILES
O=C(C)OCOC1=C(CCC(OCOC(C)=O)=O)C=C(C2(C(C=CC(C(OCOC(C)=O)=O)=C3)=C3C(O2)=O)C(C(O4)=C5)=CC(CCC(OCOC(C)=O)=O)=C5OCOC(C)=O)C4=C1.O=C(C)OCOC6=C(CCC(OCOC(C)=O)=O)C=C(C7(C(C=C(C(OCOC(C)=O)=O)C=C8)=C8C(O7)=O)C(C(O9)=C%10)=CC(CCC(OCOC(C)=O)=O)=C%10OCOC(C)=O)C9=C6
Molecular Formula
C42H40O21
Molecular Weight
880.7
Solubility
Soluble in ,Acetonitrile, DMF, DMSO
Storage
Store at -20°C, protect from light,unstable in solution, ready to use.
General tips
Select a suitable solvent according to the product solubility data. Once prepared, aliquot the stock solution to avoid repeated freeze-thaw cycles.
Use stock solutions within 6 months when stored at -80°C or within 1 month when stored at -20°C.
To improve solubility, warm the tube to 37°C and sonicate it for an appropriate period.
Use stock solutions within 6 months when stored at -80°C or within 1 month when stored at -20°C.
To improve solubility, warm the tube to 37°C and sonicate it for an appropriate period.
Shipping Condition
Evaluation samples are shipped with blue ice. Other available sizes are shipped at room temperature or with blue ice upon request.
CalculatorsMolarity / Dilution / Molecular Weight / Unit Conversion / In Vivo Formulation / Solubility
g/mol
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