Fura-2 QBT Calcium Kit
The Fura-2 QBT Calcium Kit eliminates the cause of data variability and reduces the number of steps compared to conventional wash protocols using Fura-2. Leveraging our proprietary masking technology with the industry standard Fura-2 ratiometric calcium indicator, researchers can:
- See largest Fura-2 signal window available
- Eliminate wash artifacts and increase throughput with homogeneous assay
- Minimize the impact of uneven dye loading and leakage on results
- Interrogate low-density, weakly or non-adherent cells using no-wash protocol
- Assay on FlexStation® 3 and FLIPR® Tetra Systems
- Reference thousands of citations where Fura-2 has been used for cell lines and target
Data of Fura-2 QBT Calcium Kit
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Agonism of the Muscarinic M1 Receptor on CHO M1 Cells
On the FlexStation® 3 Multi-Mode Microplate Reader, carbachol was used to stimulate the muscarinic M1 receptor on CHO M1 cells to generate concentration response curves (CRC), comparing Fura-2 QBT Dye to the BD Kit and the traditional Fura-2 wash protocol. EC50 values were similar for all methods, and were within range of published values.
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Antagonism of the Muscarinic M1 Receptor on CHO M1 Cells
On the FlexStation 3 Multi-Mode Microplate Reader, 50 nM carbachol was used as agonist challenge versus an antagonist CRC of atropine. This experiment Fura-2 QBT Calcium Kit provided the largest signal window and most robust Z-factors at EC80.
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Agonism of Endogenous H1 Receptor on HeLa Cells
Histamine was used to stimulate the endogenous histamine receptor on HeLa cells. Assay was measured using the FLIPR Tetra® High-Throughput Cellular Screening System with UV LEDs. Generated concentration response curves (CRC) comparing Fura-2 QBT Dye to the BD Ratiometric Calcium Kit and traditional Fura-2 wash protocol. EC50 values were within a half log while the Z-factor at EC80 was the largest with the Fura-2 QBT Calcium Kit.
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Antagonism of Endogenous H1 Receptor on HeLa Cells
Assay was measured using the FLIPR Tetra High-Throughput Cellular Screening System with UV LEDs and used similar experimental setup and system settings in Figure 3. 40 nM histamine was used as agonist challenge versus an antagonist CRC of pyrilamine.
Technology of Fura-2 QBT Calcium Kit
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Assay Kit Mechanism
Fura-2 QBT Calcium Kit utilizes Fura-2, AM a calcium sensitive dye that is absorbed into the cell's cytoplasm during incubation, where lipophilic blocking groups are cleaved resulting in a negatively charged fluorescent dye that stays within the cell. Upon target activation, intracellular calcium is released, binding with the dye, thereby increasing fluorescence intensity. This kit utilizes an extracellular masking technology to block background fluorescence and increase your assay's signal window, and decrease assay steps.
Calcium-Flux signal change
Calcium-bound Fura-2 is excited at 340/510 nm and the unbound form at 380/510 nm. Upon calcium-flux, the cytoplasm's calcium concentration increases, causing the fluorescence intensity at 340/510 nm to increase proportionately. In contrast, the 380/510 nm intensity declines in synchronization with the diminishing concentration of unbound dye. By ratioing fluorescence intensities produced by excitation at two wavelengths, artifacts associated with uneven dye distribution and cell number variation are minimized because they effect both measurements equally.
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Spectral Shift of Fura-2 QBT Calcium Dye When Bound to Ca2+
Fura-2 QBT™ Calcium Dye is excited at 340nm and 380 nm, with emission at 510 nm. Upon calcium flux, one can measure changes in intracellular calcium concentration.
Resources of Fura-2 QBT Calcium Kit
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
Fura-2 QBT Ratiometric Calcium Kit, a Novel Homogeneous Assay for Measuring Calcium Flux Assays on the FLIPR Tetra System and the FlexStation 3 Microplate Reader
Fura-2 QBT Ratiometric Calcium Kit, a Novel Homogeneous Assay for Measuring Calcium Flux Assays on the FLIPR Tetra System and the FlexStation 3 Microplate Reader
This PDF introduces Fura-2 QBT ratiometric calcium kit which saves time and buffer costs & offers less well to well variability, and a larger signal window.
Application Note
Fura-2 QBT Calcium Kit: A homogenous Fura-2 calcium assay
Fura-2 QBT Calcium Kit: A homogenous Fura-2 calcium assay
The Fura-2 QBT Calcium Kit from Molecular Devices incorporates proven quench based technology with a ratiometric Fura-2 calcium indicator to provide a homogenous assay to minimize cell…
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…
Compatible Products & Services of Fura-2 QBT Calcium Kit
Featured Applications
Assay Kit Mechanism
Fura-2 QBT Calcium Kit utilizes Fura-2, AM a calcium sensitive dye that is absorbed into the cell's…
Calcium-Flux signal change
Calcium-bound Fura-2 is excited at 340/510 nm and the unbound form at 380/510 nm. Upon calcium-flux…
Spectral Shift of Fura-2 QBT Calcium Dye When Bound to Ca2+
Fura-2 QBT™ Calcium Dye is excited at 340nm and 380 nm, with emission at 510 nm. Upon calcium flux…
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