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FLIPR Calcium Assay Kits

Broadest selection of calcium kits to address diversified GPCR and ion channel targets

FLIPR® Calcium Assay Kits are the platform of choice for measuring changes in intracellular calcium during drug discovery and research. Built as the most comprehensive calcium portfolio, 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.

Key Assay Features and Benefits

  • 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
  • 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.

FLIPR Calcium 3 Assay Kit
Improves signal window with unique fluorophore that enables detection of chemokines. Quench technology designed for targets that have proteins or peptides as agonists.

FLIPR Calcium Assay Kit
First homogenous no wash kit incorporating calcium sensitive fluorophore and proprietary quench technology designed for robust calcium flux assays such as the response given by agonism of the Muscarinic M1 receptor.

Patented Technology
FLIPR Assay Kits from Molecular Devices, LLC 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.

Patent Information

FLIPR Calcium Assay Kits provide a homogeneous solution for assessing GPCRs and Ion Channels

FLPR-Ca6-diagram_450px.png

Figure 1: 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.

Molecular Devices offers the broadest selection of calcium mobilization assays to address diversified biological targets. Click on the links below to view data demonstrating the benefits of each assay kit, plus links to additional posters and application notes.

  • FLIPR® Calcium 6 & 6-QF Assay Kits - Largest signal window available enabling low signal screens, probenecid- and quench-sensitive assays
  • FLIPR Calcium 5 Assay Kit - Novel ionophore demonstrates larger signal window, at a cost effective price
  • FLIPR Calcium 4 Assay Kit - Unique masking dye enhances signal to noise ratio


FLIPR Calcium 6 Assay Kit demonstrates biggest signal window
and greater flexibility over other assay kits

Ca6_Ca5_kits_in_media_350px.png
Figure 1: 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.

 

Ca6_Ca6QF_Ca5-Ca4_kits_in_buffer_350px.png
Figure 2: 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.

 

Ca6-CHOCarb-PBX_in_buffer_350px.png
Figure 3: 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.

FLIPR Calcium 5 Assay Kit demonstrates greater signal window than previous assay kits

Figure 4: 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.

 

Figure 5: 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.

 

Figure 6: 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.

 

FLIPR Calcium 4 Assay Kit novel masking dye enhances signal-to-noise ratio

flipr_assays_calcium_4_kit_fig1_350.gif
Figure 7: 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.

 

flipr_assays_calcium_4_kit_fig3_350.gif
Figure 8: 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.

 

FLIPR Calcium Assay Kits are compatible with these systems:

For more information, view the assay kits & instrument compatibility table.

Product Name
Description
Part Number
FLIPR Calcium 6 Assay Kit
Explorer Kit

Bulk Kit

Express Kit
 
1 vial = 1 plate x 10 vials + Component B Buffer

1 vial = 10 plates x 10 vials

1 vial = 50 plates x 2 vials
 
R8190

R8191

R8195
FLIPR Calcium 6-QF Assay Kit
Explorer Kit

Bulk Kit

Express Kit

1 vial = 1 plate x 10 vials + Component B Buffer

1 vial = 10 plates x 10 vials

1 vial = 50 plates x 2 vials

R8192

R8193

R8196
FLIPR Calcium 6 & 6-QF Assay Evaluation Kit 1 vial = 1 plate x 2 vials each + Component B Buffer R8194
FLIPR Calcium 5 Assay Kit
Explorer Kit

Bulk Kit

Express Kit
 

1 vial = 1 plate x 10 vials + Component B Buffer

1 vial = 10 plates x 10 vials

1 vial = 50 plates x 2 vials

R8185

R8186

R8187
FLIPR Calcium 4 Assay Kit
Explorer Kit

Bulk Kit

Express Kit

1 vial = 1 plate x 10 vials + Component B Buffer

1 vial = 10 plates x 10 vials

1 vial = 50 plates x 2 vials

R8142

R8141

R8143
FLIPR Calcium 4 Assay Kit
Explorer Kit

Bulk Kit

Express Kit

1 vial = 1 plate x 10 vials + Component B Buffer

1 vial = 10 plates x 10 vials

1 vial = 50 plates x 2 vials

R8091

R8090

R8108
FLIPR Calcium 3, 4, 5 Assay
Evaluation Kit

1 vial = 1 plate x 3 vials each + Component B Buffer

R8172
FLIPR Calcium Assay Kit
Explorer Kit

Bulk Kit

1 vial = 1 plate x 10 vials + Component B Buffer

1 vial = 10 plates x 10 vials

R8041

R8033
Component B Buffer
HBSS + Ca2+, Mg2+ and 20 mM HEPES

*Provided with Explorer and Evaluation Kits only