What Is A Flipr Assay Used For?

Basically, the FLIPR assay is used to measure changes in intracellular calcium levels in the context of drug discovery and basic research. This is done by detecting agonists, antagonists, and allosteric modulators of GPCRs. It is also used to identify ion channel targets with significant Ca 2+ permeability.

Detecting agonists, antagonists, and allosteric modulators of GPCRs

Detecting agonists, antagonists, and allosteric modulators of GPCRs has become a critical part of drug discovery. More than one third of all marketed drugs are active at GPCRs, and GPCRs are a key class of drug targets. GPCRs can be classified by their sequence homology and downstream signalling pathways.

The use of small molecule GPCR agonists is increasing as drug discovery efforts progress. GPCR agonists are often useful as research tools, and they also provide chemical starting points for drug development.

Measuring changes in intracellular calcium during drug discovery and basic research

During drug discovery and basic research, measuring changes in intracellular calcium is important for determining drug response, the mechanism of action, and side effects. Calcium is an important second messenger in cells. Calcium ions interact with proteins to alter their conformation. The ions also affect local electrostatic fields.

Calcium biosensors are used to study calcium signalling in various cell types. They are also used to study real-time kinetic readouts. Various transporters, enzymes, and pumps play a role in calcium signal generation.

Detecting ion channel targets with significant Ca 2+ permeability

Using Calcium-sensitive dyes, the FLIPR(tm) assay is used for detecting ion channel targets that exhibit significant Ca 2+ permeability. These channels are important for a variety of physiological processes. The assay requires the measurement of intracellular calcium levels and the changes in functional activity over time.

The instrument contains a cooled CCD camera imaging system and integrated pipettor capabilities. In addition to detecting allosteric modulators, it can also detect antagonists. The instrument provides low-noise detection.

Optimal dye loading for FLIPR assays

Optimal dye loading for flipr assays can be tricky business. The most optimal solution for this challenge is to purchase a dedicated instrument such as the eminence divisible Multidrop Combi or the BioRAPTR FRD, which come with pre-filled wells and a dedicated pin tool head that delivers the 20 nL/well DMSO compound solution you need.

In the interest of minimizing dilutions and maximizing fidelity, the following protocol is used: a) pre-warm the plate, b) add the fluorescent dye, c) incubate for 45 min at 37 degC, and d) cool in a controlled manner, preferably overnight.

FLIPR Penta High-Throughput Cellular Screening System

Molecular Devices FLIPR Penta is a high-throughput cellular screening line that enables high-throughput measurement of intracellular ion flux. The system can be configured based on the user’s needs and supports a range of assays, including calcium flux, membrane potential, GPCR activity, and more.

This system provides fast kinetics, simultaneous pipette and read capabilities, and exceptional luminescence performance. It can be configured with a high-speed EMCCD camera or a fluorescence camera, filters, and pipette heads.