The FLIPR Assay Model
The FLIPR assay model is one of the simplest and most useful assays that is used to measure the calcium level of various samples. It consists of two components, a Ca(2+) assay and a fluorescence assay. In this article, we will briefly describe how these two components work. Continue reading to learn more about the FLIPR assay model:
The FLIPR Ca(2+) assay is a fluorescent monitoring device which measures fluorescence emission from thousands of cells per well. It was designed to provide high throughput screening of ion channels. However, it has also contributed substantially to the pharmacological characterization of receptors.
It is a useful technique for modern drug discovery. A large number of proteins are involved in the regulation of intracellular calcium levels. In addition, calcium ions are ubiquitous second messengers in most cellular systems. They are known to modulate a wide variety of cellular processes. Therefore, they are very important targets for drug research and development. Hundreds of proteins transduce changes in intracellular calcium levels into downstream cellular signals.
CaV3.1 Fluorescence Assays
Fluorescent CaV3.1 assays can provide a reliable means of determining channel activity. However, the accuracy of the assays is limited. Here, we compared the accuracy of fluorescent-based assays with manual patch-clamp techniques. We evaluated the correlation of the two assays with the IC50 values published in the literature for CaV3.1.
We found that the CaV3.1 fluorescence assays correlated very well with the IC50 values determined by the patch-clamp assays. On average, 73% of the CaV3.2 fluorescence assay values were within threefold of the values from the patch-clamp assays.
The results of this study provide important information about the relationship between cytoskeletal interactions and T-type calcium channel trafficking. Cav3.2 deletion reduces whole cell current density, which is consistent with the role of cytoskeletal elements in channel regulation.
The FLIPR Tetra is the next-generation FLIPR. It is designed to meet the needs of advanced research professionals. Designed to expand the range of assays that can be conducted, it is also a high-throughput platform. With its industry-leading capabilities, it accelerates time to market for new products.
Molecular Devices’ FLIPR Tetra offers users a new approach to assay design. This assay platform is designed to generate high-quality biochemical kinetic data in a convenient, high-throughput format. Compared to standard readers, FLIPR offers 18-fold greater throughput.
This is achieved through a number of factors, including a high-sensitivity charge-coupled device camera that records full-plate data at a single read. In addition, the Tetra pipetting head is optimized to reduce evaporation and ensure homogeneous reaction mixtures.
Using this innovative plate reader, it is possible to assess multiple targets in a single study. The device is compatible with a wide range of assays, from fluorescent to luminescent assays. Moreover, it is also used for routine profiling efforts.