Overview of FLIPR Membrane Potential Assay Kits
Each homogeneous assay kit utilizes a proprietary indicator dye and quencher combination to maximize cell line/channel/compound applicability, while eliminating causes of data variability. This unique formula responds 10 times faster and has greater temperature stability than traditional dyes, providing high quality screening data that shows good correlation with manual patch clamp assays.
Because ion channel activity is highly sensitive and potentially impacted by subtle chemical changes, two FLIPR® Membrane Potential Assay Kits (Red and Blue) are available to select the optimal conditions for your delicate ion channel targets. Both formulations utilize Molecular Devices proprietary quench technology to enhance signal windows and yield acceptable Z-scores to screen a variety of targets, including TRP, ligand-, cyclic nucleotide- and voltage-gated channels.
Unlike traditional dyes such as DiBAC, FLIPR Membrane Potential Assay Kit detects bidirectional gradient changes so you can monitor both variable and control conditions within a single experiment. We recommend evaluating both assay kits to discern which formulation is right for your target.
Latest Resources
Data of FLIPR Membrane Potential Assay Kits
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Comparison of Patch Clamp and FLIPR Membrane Potential Assay Kit Data
The manual patch clamp method is able to detect fast responses allowing detection of very rapid changes in membrane potential. Comparing data generated using the FLIPR Membrane Potential Assay Kits with results from the manual patch clamp method shows good correlation (Figures 1 and 2). Both the opening and closing of the ion channel can be observed. This differs from DiBAC, which can only show unidirectional changes in membrane potential (Figure 3).
Comparison between patch clamp (mV) and FLIPR Membrane Potential (fluorescence) Assays on CHO cells expressing a voltage-gated K+ channel. Data courtesy of Michael Xie, Millenium Pharmaceuticals.
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Fluorescence vs Membrane Potential Change
Correlation of changes in membrane potential to fluorescence assays on the FLIPR Instrument: CHO cells transfected with K+ channel exposed to various concentrations of potassium. Data courtesy of Michael Xie, Millenium Pharmaceuticals.
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Ligand-gated Calcium Channel Comparison
Comparison between FLIPR Membrane Potential Assay Kit, DiBAC, and Fluo-3 assays on ligand-gated Ca2+ channels. Data courtesy of Michael Xie, Millenium Pharmaceuticals.
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Two Membrane Potential Kit Quench Formulations Options
Because ion channel activity is sensitive to interference, and chemical interference with a particular ion channel is highly unpredictable, the FLIPR Membrane Potential Assay Kits have two formulations. Both formulations combine the advantages of our proprietary membrane potential indicator dye with our patented quench technology. One formulation uses a blue quencher and the other formulation uses a red quencher. We recommend that both versions be evaluated for each individual target/cell line to determine which formulation will provide optimal performance.
Modulation of a Nav1.5 channel in CHL-hH1 cells by tetrodotoxin. In this assay, 30 mM veratridine is used to hold the sodium channel in its open state. Modulation occurs as tetrodotoxin concentration increases. A rapid influx of Na+ into the cell occurs, subsequently depolarizing the membrane, and leading to an increase in fluorescence. In this assay, the Membrane Potential Red Kit showed the larger signal window and larger Z factor.
Technology of FLIPR Membrane Potential Assay Kits
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Fluorescence intensity changes with increase or decrease in cellular membrane potential
FLIPR Membrane Potential Assay Kits detect ion channel modulation by increasing or decreasing the fluorescent signal as cellular membrane potential changes. The fluorescent signal increases in intensity during membrane depolarization as dye follows the positively charged ions inside the cell. During membrane hyperpolarization, fluorescent signal decreases in intensity as dye follows the positively charged ions out of the cell. The kits are uniquely suited for use with the simultaneous pipet and read capability of the FLIPR® Tetra System and FlexStation® 3 Microplate Readers to capture fast kinetics associated with ion channel activation.
The kits employ a quenching dye to reduce background fluorescence and improve the signal-to-noise ratio. The patented quench technology (U.S. patent number 6,420,183, EPO patent number 0 906 572) is offered to drug discovery and life science researchers exclusively by Molecular Devices through the purchase of FLIPR Assay Kits.
Resources of FLIPR Membrane Potential Assay Kits
eBook
Optimized Assays for Breakthrough Research
Optimized Assays for Breakthrough Research
Biology research has been greatly simplified by the availability of standardized assay kits, instruments, and assay protocols from commercial manufacturers.
Scientific Poster
FLIPR TETRA™ Membrane Potential Kit Assay Development in 384-Well and 1536 -Well Format
FLIPR TETRA™ Membrane Potential Kit Assay Development in 384-Well and 1536 -Well Format
Explore this poster to get details about FLIPR TETRA Membrane Potential Kit Assay and comparison of 384 to 1536 well format.
Data Sheet
FLIPR Membrane Potential Assay Kits
FLIPR Membrane Potential Assay Kits
Get this datasheet to learn about FLIPR Membrane Potential Assay Kits which deliver high throughput while showing good correlation with manual patch clamping data.
Application Note
Measuring Membrane Potential using the FLIPR® Membrane Potential Assay Kit on Fluorometric Imaging Plate Reader (FLIPR) Systems
Measuring Membrane Potential using the FLIPR® Membrane Potential Assay Kit on Fluorometric Imaging Plate Reader (FLIPR) Systems
Molecular Devices Corporation developed the FLIPR Membrane Potential Assay Kit to provide a fast, simple and reliable fluorescence assay for detecting changes in membrane potential.…
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Application Note
Development of a Cav 1.3 channel assay using optogenetic methods on the FLIPR Tetra System
Development of a Cav 1.3 channel assay using optogenetic methods on the FLIPR Tetra System
In this study, we demonstrate the novel utility of optogenetic tools to control membrane potential in a reversible and precise fashion for screening state-dependent calcium channel…
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Application Note
Optimization of NaV1.5 channel assay with FLIPR Membrane Potential Assay Kits
Optimization of NaV1.5 channel assay with FLIPR Membrane Potential Assay Kits
FLIPR® Membrane Potential (FMP) Assay Kits provide a rapid and reliable fluorescence-based method to detect changes in membrane potential brought about by compounds that modulate or…
Publications
A Chemical-Biological Similarity-Based Grouping of Complex Substances as a Prototype Approach for Evaluating Chemical Alternatives
A Chemical-Biological Similarity-Based Grouping of Complex Substances as a Prototype Approach for Evaluating Chemical Alternatives
Comparative assessment of potential human health impacts is a critical step in evaluating both chemical alternatives and existing products on the market. Most alternatives assessment…
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Publications
High-Content Assay Multiplexing for Toxicity Screening in Induced Pluripotent Stem Cell-Derived Cardiomyocytes and Hepatocytes
High-Content Assay Multiplexing for Toxicity Screening in Induced Pluripotent Stem Cell-Derived Cardiomyocytes and Hepatocytes
Cell-based high-content screening (HCS) assays have become an increasingly attractive alternative to traditional in vitro and in vivo testing in pharmaceutical drug development and t…
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Publications
Assessment of Beating Paramaters in Human Induced Pluripotent Stem Cells Enables Quantitative in Vitro Screening for Cadiotoxicity
Assessment of Beating Paramaters in Human Induced Pluripotent Stem Cells Enables Quantitative in Vitro Screening for Cadiotoxicity
Human induced pluripotent stem cell (iPSC)-derived cardiomyocytes show promise for screening during early drug development. Here, we tested a hypothesis that in vitro assessment of m…
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Multiparameter In Vitro Assessment of Compound Effects on Cardiomyocyte Physiology Using iPSC Cells
Multiparameter In Vitro Assessment of Compound Effects on Cardiomyocyte Physiology Using iPSC Cells
A large percentage of drugs fail in clinical studies due to cardiac toxicity; thus, development of sensitive in vitro assays that can evaluate potential adverse effects on cardiomyoc…
