FIVEPHOTON MEMBRANE POTENTIAL ION CHANNEL ASSAY KIT | Fluorescent Voltage Sensitive Dye Kit (kw. Voltage-sensitive probes: Part MPF-Kit)

• 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 and prokaryotic cell types as well as tissues and vessels due to the lipophilic nature of the dye mixtures.
• Generates highly reproducible and reliable data for drug screening applications and publication.
• Economically priced:  Each kit is applicable for multiple 96-well dishes. 
  
• Use bottom-reading fluorescent plate reader to measure signal.
• Independently validated by our customers.

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) primarily due to Na⁺, K⁺, and Cl^- ion concentrations gradients which are maintained by active transport.  Voltage sensitive probes (VSP) offer a convenient method to measure translocation of ions across cellular membranes. The FIVEphoton Biochemicals membrane potential assay kits provide a unique formulation of oxonol dyes and quencher.  The non-membrane permeant quencher is a component of the mixture that eliminates the need for washes and minimizes data scatter.

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.

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Representative Measurements

 

Figure legend.  Dose response curves generated with the Fivephoton Biochemicals Membrane Potential Assay Kit to standard sodium channel blockers in transfected CHO cells.  Fluorescent emission was measured with a fluorescent plate reader.  CHO cells were over expressing the TTX sensitive NaV channel subunit.  Veratridine at its EC80 (i.g. 100uM) was added to open NaV channels.  NaV channel blockers that were applied include the following:  Amitriptyline, Dibucaine, Tetracaine, Tetrodotoxin (TTX).  Data with respect to control was standardized on fluoresent emission intensity measurements.

 

 

 

 

 

 

 

 

 

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Ion Channel Membrane Potential Assay Kits

 

               Kits with pre-validated dye-quencher mix

             

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

 

       Use available options menu to order.  Bulk quantities are available:  Please inquire.

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. 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.
6. 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.
7. 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.
8. 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..
9. 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.
10. 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.
11. 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.
12. 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.

kw.  membrane potential dye; membrane potential probe; voltage sensitive dye; voltage sensitive probes; fluorescent dye/probe; membrane voltage dye/probe, 

ion channel assay,  voltage sensitive probes, voltage sensitive dye, membrane potential dye, potassium channel, chloride channel, sodium channel, ligand-gated ion channel, TRP, ABC transporter, CFTR, K+ channel, Cl- channel, Na+ channel, cell-based assay