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2025.03.04 18:54

[Five Photon 공식 대리점]Fivephoton Membrane Potential Dye Ion Channel Assay Kit(MPF-Kit)

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Fivephoton Membrane Potential Dye Ion Channel Assay Kit


Ion Channel Assay Kit Using Fluorescent Voltage-Sensitive Dye-Quencher Mixtures.  Applicable to Eurkaryotic and Prokaryotic Cells, Tissues, and Vessels
 
 
Lead time:  Approx 1 week
 
Highlights of the Fivephoton Biochemicals Fluorescent Membrane Potential Dye Ion Channel Assay 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.

View Protocol


Ion Channel Membrane Potential Dye 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 in eukaryotic and prokaryotic cells.  The kit is designed to detect ion flux through the majority of ion 
channels, transporters and receptors.  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.  
 
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.

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.

 
 












 
 


 
 
 
 
 
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.
 
Product References
 

Eurkayotes

1.  Thompson, Mackenzie J., et al. (2022). "Distinct functional roles for the M4 α-helix from each homologous subunit in the hetero-pentameric ligand-gated ion channel nAChR." Journal of Biological Chemistry. Volume 298, Issue 7, July 2022, 102104.  Link to article

2.  Purnell, Marcy C., and Terence J. Skrinjar. "Bioelectric Field Enhancement: The Influence on Membrane Potential and Cell Migration In Vitro." Advances in Wound Care 5.12 (2016): 539-545.  Link to article

3.  Igor M Pongrac, Ivan Pavičić, Mirta Milić, Lada Brkić Ahmed, Michal Babič, Daniel Horák, Ivana Vinković Vrček, Srećko Gajović.  2016.  Oxidative stress response in 

neural stem cells exposed to different superparamagnetic iron oxide nanoparticles.  International Journal of Nanomedicine.  Volume 2016:11 Pages 1701—1715.  Link to article

4.  Binepal, G., Gill, K., Crowley, P., Cordova, M., Brady, L. J., Senadheera, D. B., & Cvitkovitch, D. G. (2016). The Trk2 potassium transport system in Streptococcus 
mutans and its role in potassium homeostasis, biofilm formation and stress tolerance. Journal of Bacteriology, JB-00813.  Link to article
 
Bacteria
 
1.  Singh, Kamna, et al. "The copYAZ operon functions in copper efflux, biofilm formation, genetic transformation, and stress tolerance in Streptococcus mutans." Journal 
of bacteriology 197.15 (2015): 2545-2557.  Link to article 

Related Products and Supplements for Ion Channel Membrane Potential Assay

Membrane Potential Measurement Kit for HTS Pharmaceutical
Screening:  click here   
                                                     
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 
  2. nerve cell in the rat cortical slice. Biol Bull 193: 261, 1997.
  3. 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.
  4. 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.
  5. 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.
  6. 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.
  7. 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.
  8. 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.
  9. 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..
  10. 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.
  11. 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.
  12. 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.
  13. 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.

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