BCECF AM Documentation

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BCECF AM Product Specifications

Fluorescence Spectrum

Molecular Weight

821 g/mol

Solubility

DMSO

pKa

7

Odor

None

Toxicology Data

Not known

Shelf Life

Valid for one year after delivery, if stored properly

TLC

Solvent

Hexane/ Ethyl Acetate 2:1

Rf

0.75

HPLC

Column

C18

Detector Settings

254 nm, 469 nm

Purity

Four peaks corresponding to the four components

Absorbance Spectrum

Solvent

Methanol

Absorbance max

469 ± 3 nm

ε

21,000 M-1cm-1

After hydrolysis with KOH

Excitation max

500/450 nm hi/lo pH

Emission max

531 nm

1H NMR

All relevant peaks present

Solvent

Deuterated Chloroform

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BCECF AM Background

Ion Indicators offers BCECF AM for two functions- as a pH indicator and as an indicator for measuring cell death and viability kinetically. The indicator was invented by Roger Tsien [Journal of Call Biology 95; 189 (1982)] as a pH indicator with an ideal pK a of 6.98 for the cell physiology pH of 7. The AM form loads into cells non-invasively and are cleaved by only living cells which have non-specific esterases to give the green-emitting dye for study. Dead cells lack esterases and can therefore be distinguished from live cells. The pK a allows for measurement of small changes in physiological pH. This indicator has been around for a long time and therefore we have added a list of references below.

  • DeClerck LS, Bridts CH, Mertens AM, Moens MM, Stevens,WJ. Use of fluorescent dyes in the determination of adherence of human leucocytes to endothelial cells and the effect of fluorochromes on cellular function. J. Immunol. Meth. 1994;172: 115–24.
  • P. Ozkan and R. Mutharasan, A Rapid Method for Measuring Intracellular pH using BCECF-AM, Biochim. Biophys. Acta, 1572:143, 2002.
  • Obexer et al., 1995, W Obexer, C Schmid, R Brun, A novel in vitro screening assay for trypanocidal activity using the fluorescent dye BCECF-AM, Trop. Med. Parasitol., 46 (1995), pp. 45-48
  • Franck, P., Petitipain, N., Cherlet, M., Dardennes, M., Maachi, F., Schutz, B., Poisson, L. & Nabet, P. (1996), Measurement of intracellular pH in cultured cells by flow cytometry with BCECF-AM, J. Biotechnol. 46, 187–195.

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