FIVEphoton Membrane Potential Assay Kit (part MPFKit1)

Manufacturer Fivephoton
Category Products / Kits
Type Kit
Specific against other
Amount I kit for 5 ml assay buffer
ArtNr MPFKit1
Eclass 6.1 32161000
Eclass 9.0 32161090
Available
Highlights
Enables the cell biologist, neurobiologist and ion channel physiologist to accurately and reproducibly measure changes in ion flux without patch clamp technique.
Broadly applicable to most eukaryotic cell types.
Rapid response times using a proprietary mixtures of fluorescent indicator dyes and quenchers.
Generates highly reproducible and reliable data for publication and drug screening applications.
Economically priced. Each kit is applicable for multiple 96-well dishes.
Easily measured by most bottom-reading fluorescent plate readers.
Background
Ion Channel Membrane Potential Assay Kit

The Fivephoton Ion Channel Membrane Potential Assay Kit provides an easy-to-use, highly sensitive, accurate quantitative method to measure changes in ion flux and the cellular membrane potential using fluorescent voltage sensitive dyes. The kit is designed to detect ion flux through the majority of ion channels, transporters and receptors that give rise to membrane polarization changes. The assay offers a rapid response time, does not require washing, operates at room and physiological temperatures and provides reproducible and reliable data that correlate with the rapid opening and closing of ion channels.

The kits contain fluorescent voltage sensitive dye and quencher mixtures with spectral properties easily measured by commonly employed fluorescent plate readers. In addition to offering kits with one pre-validated dye-quencher mixture, we offer several distinct formulations of voltage sensitive dye to quencher, enabling the researcher to empirically identify the optimal dye-quencher mixture for their cell type.

Introduction to Membrane Potential Assay Dyes

The bilipid membrane of a cell typically has a transmembrane potential of approximately -60 to -80 mV (negative inside) as a consequence of mostly Na+, K+, and Cl- ion concentrations gradients which are maintained by active ion transporters. Voltage sensitive probes (VSP) offer a convenient method of detecting the translocation of these ions across cellular membranes. The thiobarbituric and bis-barbituric acid oxonols, referred to as DiSBAC2 and DiBAC3 dyes respectively, form a family of VSPs with distinct excitation parameters. The FIVEphoton Biochemicals membrane potential assay kits utilize a dye formulation which is very similar to the most popular oxonol dyes for transmembrane potential measurement.

Membrane potential assay dyes enter depolarized cells and bind to intracellular proteins or membranes, resulting in enhanced fluorescence and a red spectral shift. Increase in depolarization leads to elevated influx of voltage senstivie dye and an increase in fluorescence that can be measured by optical detectors such as fluorescent microplate readers, fluorometers and flow cytometers.
References
Antic S, Major G, Chen WR, Wuskel J, Loew L, and Zecevic D. Fast voltage-sensitive dye recording of membrane potential changes at multiple sites on an individual nerve cell in the rat cortical slice. Biol Bull 193: 261, 1997.

Baxter DF, Kirk M, Garcia AF, Raimondi A, Holmqvist MH, Flint KK, Bojanic D, Distefano PS, Curtis R, and Xie Y. A novel membrane potential-sensitive fluorescent dye improves cell-based assays for ion channels. J Biomol Screen 7: 79-85, 2002.

Beck JC and Sacktor B. Membrane potential-sensitive fluorescence changes during Na+-dependent D-glucose transport in renal brush border membrane vesicles. J Biol Chem 253: 7158-7162, 1978.

Benjamin ER, Skelton J, Hanway D, Olanrewaju S, Pruthi F, Ilyin VI, Lavery D, Victory SF, and Valenzano KJ. Validation of a fluorescent imaging plate reader membrane potential assay for high-throughput screening of glycine transporter modulators. J Biomol Screen 10: 365-373, 2005.

Coclet-Ninin J, Rochat T, Poitry S, Chanson M. Discrimination between cystic fibrosis and CFTR-corrected epithelial cells by a membrane potential-sensitive probe. Exp Lung Res 28: 181-99, 2002.

Gao LJ, Yang WD, and Liu JS. [A fluorescent dye method based on changes in membrane potential for detecting PSP toxins in shellfish]. Guang Pu Xue Yu Guang Pu Fen Xi 29: 1032-1035, 2009.

Gaskova D, Brodska B, Herman P, Vecer J, Malinsky J, Sigler K, Benada O, and Plasek J. Fluorescent probing of membrane potential in walled cells: diS-C3(3) assay in Saccharomyces cerevisiae. Yeast 14: 1189-1197, 1998.

Huang CJ, Harootunian A, Maher MP, Quan C, Raj CD, McCormack K, Numann R, Negulescu PA, and Gonzalez JE. Characterization of voltage-gated sodium-channel blockers by electrical stimulation and fluorescence detection of membrane potential. Nat Biotechnol 24: 439-446, 2006..

Mao C and Kisaalita WS. Determination of resting membrane potential of individual neuroblastoma cells (IMR-32) using a potentiometric dye (TMRM) and confocal microscopy. J Fluoresc 14: 739-743, 2004.

Matsumoto A, Doi T, Asako M, Yang SM, and Yamashita T. Optical recording of membrane potential on isolated spiral ganglion cells of newborn mice using a voltage-sensitive dye. Acta Otolaryngol Suppl 539: 34-39, 1998.

Sguilla FS, Tedesco AC, and Bendhack LM. A membrane potential-sensitive dye for vascular smooth muscle cells assays. Biochem Biophys Res Commun 301: 113-118, 2003.

Tominaga Y, Ichikawa M, and Tominaga T. Membrane potential response profiles of CA1 pyramidal cells probed with voltage-sensitive dye optical imaging in rat hippocampal slices reveal the impact of GABA(A)-mediated feed-forward inhibition in signal propagation. Neurosci Res 64: 152-161, 2009.
Amount: I kit for 5 ml assay buffer
Available: In stock
Listprice: €334.85
Price: €334.85
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