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FLIPR Calcium Assay Kits
FLIPR® Calcium Assay Kits are the platform of choice for measuring changes of intracellular calcium for drug discovery and basic research.
- Maximizes fluorescent signal from a wide range of cell types by reducing cell wash artifacts
- Reduces well-to-well variability, improving assay quality (Z' factor) and reliability (CV %) of high-throughput screens
FLIPR Calcium Assay Kit
Built as the most comprehensive portfolio of calcium reagents, the kits deliver pre-optimized, homogeneous, fluorescence-based formulations to expedite assay development and screening of GPCR and ion channel targets. FLIPR Calcium Assay Kits provide the most comprehensive method for detecting intracellular calcium changes in a simple and reliable homogeneous assay format. Select from six formulations to complement your target of interest.
FLIPR Assay Kits employ a quenching dye to reduce background fluorescence and improve the signal-to-noise ratio. The patented quench technology is offered to drug discovery and life science researchers exclusively by Molecular Devices, through the purchase of FLIPR Assay Kits.
Key Assay Features and Benefits
- Universal mix-and-read protocol accelerates assay workflow and increases throughput
- Superior signal-to-noise ratio facilitates confirmation of endogenous or transiently transfected receptor activity during assay development
- Pre-optimized and validated protocols for the FLIPR® Tetra and FlexStation® Systems ensure you can navigate both routine and unconventional cell lines and targets
FLIPR Calcium 6 and 6-QF Assay Kits
This exclusive fluorophore features the highest quantum yield of any calcium indicator on the market, delivering the greatest signal window available in a calcium assay kit. The substantial increase in signal amplitude helps to rekindle low signal screens; including endogenous, primary or stem cell targets. Additionally, the novel dye is unaffected by organic anion exchange proteins, enabling measurement with minimal to no probenecid present.
When utilized in the quench-free (QF) formulation, the dye is ideal for quench sensitive targets or assays requiring multiplexing of calcium-flux and luminescent assays within one well.
FLIPR Calcium 5 Assay Kit
A novel fluorophore that amplifies the assay signal window while leveraging the superior masking technology used in the FLIPR Calcium 4 Assay Kit to reduce background fluorescence. In addition to the above assays, receptors with low receptor expression and other targets that give a small calcium response are candidates for this kit.
FLIPR Calcium 4 Assay Kit
Contains a revised proprietary masking dye with higher light extinction in the extracellular solution to significantly reduce background and increase signal window. It is more inert and reduces compound interference. Successful targets include calcium channels, chemokine receptors, and transiently transfected receptors.
Latest Resources
Data of FLIPR Calcium Assay Kits
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FLIPR Calcium 6 Assay Kit signal window
Comparison of fluorescent signal response to calcium flux during carbachol agonism of Muscarinic M1 receptor in CHO M1 cells. This illustrates the signal-to-noise ratios offered by homogenous assay run in media using the FLIPR Calcium 6 Assay Kit. Combining a novel fluorophore and the proven quench technology, the FLIPR Calcium 6 Assay Kit shows the highest signal to background ratio.
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Quencher free signal window
When performing no-wash assays (initial media removal, replaced with assay buffer, no-wash after dye-loading) the novel fluorophore of FLIPR Calcium 6 Assay Kits demonstrates highest signal to background ratio. Obtain same high quality performance in presence or absence of masking dye.
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Probenecid free signal window
Novel fluorophore is more resistant to organic anion exchange proteins found in some cell lines. No signal seen when using FLIPR Calcium 5 Assay Kit without probenecid (Ca5 Kit no PBX). With FLIPR Calcium 6 Assay Kit (Ca6), the signal is smaller without probenecid, however EC50s are conserved and Z' still greater than 0.5.
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FLIPR Calcium 5, 4, & 3 Assay Kit comparison
Comparison of fluorescent signal response to calcium flux during acetylcholine agonism of Muscarinic M1 receptor in CHO M1 cells. This illustrates the signal-to-noise ratios offered by various FLIPR Calcium Assay Kits with quench technology. Combining the novel fluorophore and the proven quench technology of the FLIPR Calcium 4 Assay Kit, the FLIPR Calcium 5 Assay Kit shows higher signal to background ratio than prior kits.
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Antagonism data with FLIPR Calcium 5 Assay Kit
Antagonism of calcium flux in response to EC80 acetylcholine in CHO M1 cells evaluated with the FLIPR Calcium 5 Assay Kit and the FLIPR Tetra System, demonstrating a large signal window and performance consistent with previously published data.
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Signal trace comparison
Signal trace on the FLIPR Tetra System of ATP agonism of PY receptor in HEK 293 cells compared using different calcium mobilization kits. Consistent EC50 values among all kits were recorded with no increase in background. The signal window was substantially larger for the FLIPR Calcium 5 Assay Kit.
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FLIPR Calcium 4 & 3 Assay Kit comparison
Reduced background in the FLIPR Calcium 4 Assay Kit (orange) leads to improved ΔF/Fo values compared to the FLIPR Calcium 3 Assay Kit (green) in an assay with especially small calcium response. Assay: Endogenous response to carbachol in HEK 293 cells in a 384-well plate, 10,000 cells/well, media removal before assay on the FLIPR Tetra System.
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FLIPR Calcium 4 and 3 Assay Kits vs competitor
In many cases FLIPR Calcium 4 and Calcium 3 Assay Kits perform equally well while at the same time outperforming competitive kits as shown by a Molecular Devices customer in the data above. Assay: Undisclosed non-adherent cells stimulated with a protein activator, no media removal, assay was done on a FLIPR 3 instrument.
Technology of FLIPR Calcium Assay Kits
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FLIPR Calcium Assay Kits Provide a Homogeneous Solution for Assessing GPCRs and Ion Channels
FLIPR Calcium Assay Kits utilize a calcium sensitive dye that is absorbed into the cell's cytoplasm during incubation. Upon ligand-receptor binding, intracellular calcium is released, binding with the dye, thereby increasing fluorescence intensity. All FLIPR Calcium Assay Kits utilize an extracellular masking technology to block background fluorescence and increase your assay's signal window. The FLIPR Calcium 6-QF Assay Kit is the one exception, enabling assay performance without a masking technology, making it an ideal choice for quench-sensitive applications such as multiplexing with luminescent indicators.
Ordering Options of FLIPR Calcium Assay Kits
FLIPR Calcium 6 Assay Kit
Evaluation Kit (1 Plate) - PN: R8194
- (3) vials of component A* for Calcium 6
- (3) vials of component A for Calcium 6-QF
- (1) bottle of dilution buffer (Component B)
- (3) vials of component C
* Each reagent vial (Component A, C) is sufficient for 1 plate (96-, 384-, 1536-well). Each kit is sufficient for 3 Calcium 6 assay plates and 3 Calcium 6-QF assay plates.
#R8194
Explorer Kit (10 Plates) - PN: R8190
- (10) vials of component A*
- (1) bottle of dilution buffer (Component B)
* Each reagent vial (Component A) is sufficient for 1 plate (96-, 384-, 1536-well). Each kit is sufficient for 10 plates.
#R8190
Bulk Kit (100 Plates) - PN: R8191
- Component A (10 vials*)
* Each vial is sufficient for 10 plates (96- and 384-well), or 7 plates (1536-well).
The entire Bulk Kit is sufficient for 100 plates (96- and 384-well), or 70 plates (1536-well).
#R8191
Express Kit (100 Plates) - PN: R8195
- Component A (2 vials*)
* Each vial is sufficient for 50 plates (96- and 384-well), or 35 plates (1536-well).
The entire Express Kit is sufficient for 100 plates (96- and 384-well), or 70 plates (1536-well).
#R8195
FLIPR Calcium 6-QF Assay Kit
Explorer Kit - PN: R8192
- (10) vials of component A*
- (1) bottle of dilution buffer (Component B)
- (10) vials of component C
* Each reagent vial (Component A, C) is sufficient for 1 plate (96-, 384-, 1536-well). Each kit is sufficient for 10 plates.
#R8192
Bulk Kit - PN: R8193
- (10) vials of component A*
- (10) vials of component C
* Each reagent vial (Component A, C) is sufficient for 10 plates (96-, 384-, 1536-well). Each kit is sufficient for 100 plates.
#R8193
Express Kit - PN: R8196
- (2) vials of component A*
- (2) vials of component C
* Each reagent vial (Component A, C) is sufficient for 50 plates (96-, 384-, 1536-well). Each kit is sufficient for 100 plates.
#R8196
FLIPR Calcium 5 Assay Kit
Explorer Kit - PN: R8185
- (10) vials of component A*
- (1) bottle of dilution buffer (Component B)
* Each reagent vial (Component A) is sufficient for 1 plate (96-, 384-, 1536-well). Each kit is sufficient for 10 plates.
#R8185
Bulk Kit - PN: R8186
- (10) vials of component A*
* Each reagent vial (Component A) is sufficient for 10 plates (96-, 384-, 1536-well). Each kit is sufficient for 100 plates.
#R8186
Express Kit - PN: R8187
- (2) vials of component A*
* Each reagent vial (Component A) is sufficient for 50 plates (96-, 384-, 1536-well). Each kit is sufficient for 100 plates.
#R8187
FLIPR Calcium 4 Assay Kit
Explorer Kit - PN: R8142
- (10) vials of component A*
- (1) bottle of dilution buffer (Component B)
* Each reagent vial (Component A) is sufficient for 1 plate (96-, 384-, 1536-well). Each kit is sufficient for 10 plates.
#R8142
Bulk Kit - PN: R8141
- (10) vials of component A*
* Each reagent vial (Component A) is sufficient for 10 plates (96-, 384-, 1536-well). Each kit is sufficient for 100 plates.
#R8141
Express Kit - PN: R8143
- (2) vials of component A*
* Each reagent vial (Component A) is sufficient for 50 plates (96-, 384-, 1536-well). Each kit is sufficient for 100 plates.
#R8143
Resources of FLIPR Calcium Assay Kits
Application Note
Automated Monitoring of Development and Activity Analysis of iPSC-derived 3D Cerebral Organoids
Automated Monitoring of Development and Activity Analysis of iPSC-derived 3D Cerebral Organoids
The human brain is highly complex which makes it challenging to study both in vitro and in-model organisms. Cultured neurons do not sufficiently recapitulate the three-dimensional (3D…
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
Validation of TRPV1 and ASIC1 Ligand-Gated Ion Channels using Automated Patch Clamp and FLIPR with novel Ca2+ detection dye
Validation of TRPV1 and ASIC1 Ligand-Gated Ion Channels using Automated Patch Clamp and FLIPR with novel Ca2+ detection dye
In one series of experiments, we constructed and validated cell based assays in which the FLIPR Calcium 6 assay kit containing this novel calcium fluorophore was used to examine the…
Scientific Poster
Optimizing a Transient Receptor Potential (TRP) Channel Assay on the FLIPRTETRA® Instrument using hTRPV1-HEK293 Cells and the FLIPR® Calcium 5 Assay Kit
Optimizing a Transient Receptor Potential (TRP) Channel Assay on the FLIPRTETRA® Instrument using hTRPV1-HEK293 Cells and the FLIPR® Calcium 5 Assay Kit
This PDF describes optimization of the TRP (transient receptor potential) channel assay on the FLIPR TETRA instrument using hTRPV1-HEK293 cells and the FLIPR calcium 5 assay kit.
Scientific Poster
Functional Characterisation of Recombinant Muscarinic M3 Receptors in Assay-Ready Frozen Cells using a Novel Calcium Assay Kit on the FLIPRTETRA® Instrument
Functional Characterisation of Recombinant Muscarinic M3 Receptors in Assay-Ready Frozen Cells using a Novel Calcium Assay Kit on the FLIPRTETRA® Instrument
This poster compares the performance of cryopreserved "frozen" cells vs. cells in culture when used for screening GPRC drug candidates on the FLIPR Tetra instrument.
Scientific Poster
Cell Based Assays on the FLIPR TETRA® System : Comparison of the FLIPR Calcium 5 Assay Kit to Other Fluorescence Based Calcium Flux Assays
Cell Based Assays on the FLIPR TETRA® System : Comparison of the FLIPR Calcium 5 Assay Kit to Other Fluorescence Based Calcium Flux Assays
This poster explores FLIPR Calcium 5 Assay Kit containing a superior performance calcium-sensitive indicator that does not require media removal & utilizes same proven quench tec…
Application Note
Enhance your calcium screens with FLIPR Calcium 6 Assay Kits
Enhance your calcium screens with FLIPR Calcium 6 Assay Kits
FLIPR Calcium Assay Kits from Molecular Devices employ sensitive calcium indicators and proprietary masking dyes to enable researchers to conduct highly sensitive fluorescent screens…
Application Note
Calcium signaling with FLIPR Calcium 6 and 6-QF Assay Kits on the FlexStation 3 reader
Calcium signaling with FLIPR Calcium 6 and 6-QF Assay Kits on the FlexStation 3 reader
FLIPR® Calcium Assay Kits from Molecular Devices employ sensitive calcium indicators and proprietary masking dyes to enable researchers to conduct highly sensitive fluorescent screens…
Application Note
Homogeneous solution for GPCR assays with FLIPR Calcium 5 Assay Kit
Homogeneous solution for GPCR assays with FLIPR Calcium 5 Assay Kit
Combining a novel fluorophore and proven masking technology, the FLIPR Calcium 5 Assay Kit delivers reliable pharmacology, a larger signal window, and improved assay performance. With…
Application Note
High throughput cardiotoxicity assays using stem cell-derived cardiomyocytes
High throughput cardiotoxicity assays using stem cell-derived cardiomyocytes
Eliminate cardiotoxic compounds and identify potential cardiac drug candidates earlier in the drug discovery process with the FLIPR Tetra System, together with ScreenWorks Peak Pro…
Application Note
Comparison of intracellular calcium measurements with FLIPR Calcium Assay Kit
Comparison of intracellular calcium measurements with FLIPR Calcium Assay Kit
The FLIPR Tetra System delivers a screening platform to address real-time kinetic cell-based assays in 96-, 384- and 1536-well format. Regardless of the FLIPR Tetra System microplate…
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…
Application Note
Monitoring Gq protein-coupled receptor activation on the SpectraMax i3x reader with injector module
Monitoring Gq protein-coupled receptor activation on the SpectraMax i3x reader with injector module
Gq protein-coupled receptor activation is commonly monitored in live cells in real time using calcium-sensitive dyes on a fluorescence plate reader. Automated liquid handling within…
Scientific Poster
Calcium Release-Activated Channel (CRAC) Assays on the FLIPR® Tetra System: Evaluation of a Novel Fluorescent Calcium Reagent
Calcium Release-Activated Channel (CRAC) Assays on the FLIPR® Tetra System: Evaluation of a Novel Fluorescent Calcium Reagent
Get detailed information about Calcium release-activated calcium channels by exploring this PDF. Learn how CRAC channels are playing an important role in intracellular Ca2+ homeostas…
Scientific Poster
Cell Based Assays on the FLIPR® Tetra System: Comparison of a Novel FLIPR® Calcium Assay to other Fluorescence based Calcium Flux Assays
Cell Based Assays on the FLIPR® Tetra System: Comparison of a Novel FLIPR® Calcium Assay to other Fluorescence based Calcium Flux Assays
Cell based assays on FLIPR Tetra System to compare a novel FLIPRCcalcium 6 and 6-QF assay to other fluorescence-based calcium flux assays.
Data Sheet
Calcium assays for the FLIPR System
Calcium assays for the FLIPR System
The FLIPR® Calcium Assay Kits have been the premier choice for measuring changes in intracellular calcium during drug discovery and research. Built as the most comprehensive calcium…
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…
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…
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…
Publications
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…


Optogenetics and the FLIPR: How to Use Light to Set up a Cav1.3 High-Throughput Screening Assay
Compatible Products & Services of FLIPR Calcium Assay Kits
How can we help advance your next big discovery?
Our highly-qualified teams are on the frontlines with our customers, conducting remote or on-site product demonstrations, webinars, and more to help you solve your tough research challenges. How can we help you today?
I’d like to…
How can we help advance your next big discovery?
Our highly-qualified teams are on the frontlines with our customers, conducting remote or on-site product demonstrations, webinars, and more to help you solve your tough research challenges. How can we help you today?
I’d like to…