FIVEPHOTON FLUORESCENT MEMBRANE POTENTIAL ASSAY KIT - High Throughput Format (Part. No. mpHTS-Kit)

Fluorescent Voltage-Sensitive Dye Membrane Potential Assay Kits   

Highlights of the Membrane Potential Assay System:

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View pull-down menu above to select option.

Part	Vials provided	Dye volume (µl) per vial	Dye concentration in vials			Cost
MPF Kit1	3	100	500X			$490
MPF Kit5	1	100	500X			$390
MPF Kit6	1	200	500X			$550
MPF Kit7	1	300	500x			$600

               

 

            Bulk Pharmacological Screen Format

Option	96-well plates per kit	Number of vials	Cost
mpHTS-KitA	50	1 (one dye-quencher mix)	$1563
mpHTS-KitB	50	6 (six dye-quencher mixes*)	$1563

              

                 *  Part mpHTS-KitB;  dye-quencher mix suitable for 8 plates per vial. 

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Introduction to Ion Channel 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, are VSPs with distinct excitation and emission 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 dyes enter depolarized cells and associate with intracellular proteins or membranes, causing enhanced fluorescence and a spectral shift to red wavelengths.  Increase in membrane depolarization leads to further influx of dye and thus an increase in fluorescence that can be measured by fluorescence spectroscopy and fluorescent plate readers.

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Representative References On Ion Channel Membrane Potential Assays

1. 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.
2. 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.
3. 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.
4. 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.
5. 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.
6. 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.
7. 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..
8. 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.
9. 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.
10. 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.
11. 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.
12. Waggoner AS. Dye indicators of membrane potential. Annu Rev Biophys Bioeng 8: 47-68, 1979.
13. Whiteaker KL, Gopalakrishnan SM, Groebe D, Shieh CC, Warrior U, Burns DJ, Coghlan MJ, Scott VE, and Gopalakrishnan M. Validation of FLIPR membrane potential dye for high throughput screening of potassium channel modulators. J Biomol Screen 6: 305-312, 2001.

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