Uses
Calcein-AM is a hydrophobic non-fluorescent probe that can permeate the plasma membrane and can be hydrolyzed to Calcein (sc-202090), which is an extremely fluorescent and negatively charged molecule. Calcein-AM can be used to stain living cells and in cytoplasmic Schwann cells has been observed to fluoresce bright green. Calcein-AM has been reported to be a neutral substrate for P-glycoprotein (Pgp) and multidrug resistance protein (MRP) and has been used in flow cytometry studies to analyze the function of P-gp and MRP.
Definition
ChEBI: Calcein am is an organic heteropentacyclic compound that is calcein in which all four carboxy group hydrogens have been substituted by (acetyloxy)methoxy groups and the hyrodgens of the two hydroxy groups have been substituted by acetyl groups. It is a a non-fluorescent probe cleaved to a fluorescent probe by non-specific intracellular esterases. It has a role as a fluorochrome. It is an acetate ester, an organic heteropentacyclic compound, a gamma-lactone, an oxaspiro compound, a member of 2-benzofurans and a xanthene dye. It is functionally related to a calcein.
General Description
Liposomes contain a self-quenching fluorescent dye called calcein. It is a fluorescent dye with excitation and emission wavelengths of 495/515 nm, respectively. It self-quenches at concentrations above 70mM and is commonly used as an indicator of lipid vesicle leakage. The leakage rate is calculated on the basis of the fluorescence increment as the entrapped calcein is leaked out of the liposomal compartment. It is used as a complexometric indicator for titration of calcium ions with EDTA, and for fluorometric determination of calcium.
References
[1] JAMES HYNES . Fluorescence-Based Cell Viability Screening Assays Using Water-Soluble Oxygen Probes[J]. SLAS Discovery, 2003, 8 3: Pages 264-272. DOI:10.1177/1087057103008003004.
[2] SABNIS R W. Handbook of Biological Dyes and Stains: Synthesis and Industrial Applications[C]. 2010: 0. DOI:10.1002/9780470586242.
[3] Polli, J.W., Wring, S.A., Humphreys, J.E., et al. Rational use of in vitro P-glycoprotein assays in drug discovery. J. Pharmacol. Exp. Ther. 299(2), 620-628 (2001).
[4] Tiberghien, F., and Loor, F. Ranking of P-glycoprotein substates and inhibitors by a calcein-AM fluorometry screening assay. Anticancer Drugs 7(5), 568-578 (1996).
[5] Szakács, G., Jakab, K., Antal, F., et al. Diagnostics of multidrug resistance in cancer. Pathol.Oncol.Res. 4(4), 251-257 (2016).
[6] X M WANG. A new microcellular cytotoxicity test based on calcein AM release.[J]. Human Immunology, 1993, 37 4: 264-270. DOI:10.1016/0198-8859(93)90510-8.
[7] ZSOLT HOLLó . Calcein accumulation as a fluorometric functional assay of the multidrug transporter[J]. Biochimica et biophysica acta. Biomembranes, 1994, 1191 2: Pages 384-388. DOI:
10.1016/0005-2736(94)90190-2.