Calcium Flux Assays

Automated platforms for conducting calcium flux assays

Calcium flux assays are fluorescence-based assays designed to detect the transient release of intracellular calcium stores. These assays typically rely on cell-permeable dyes with fluorescence properties that are calcium sensitive.

The application solutions optimized on Agilent microplate readers and automated imaging systems provide sensitive and reliable detection of calcium flux across a broad range of reagents. Integrated hardware and Agilent BioTek Gen5 software features enable the detection and relative quantification of calcium release following agonist addition within high-density microplate formats.

Dual masking with nuclear stain for subcellular characterization and percent responder studies

Gaining deeper insight into receptor-mediated responses with subcellular analysis

The addition of an indiscriminate nuclear marker (e.g., Hoechst) facilitates dual masking of cells within a population, with a primary mask placed around the nuclei and a secondary mask placed around the cell perimeter. This provides a convenient way to determine the percentage of cells that respond to treatment, as well as to measure the relative response with subcellular resolution (i.e., nuclear versus cytoplasmic signaling).

Characterizing Calcium Mobilization Using Kinetic Live Cell Imaging

Ca2+ acts as an important second messenger in diverse signaling pathways, including G protein-coupled receptors. Characterizing these pathways requires the ability to detect rapid changes in intracellular Ca2+ levels with high temporal resolution. This application note describes a live cell imaging-based approach to quantify Ca2+ flux kinetics using an Agilent BioTek Lionheart FX and Fluo-4 Ca2+ indicator dye that delivers sub-second resolution and a large assay window.

Read app note

Automated Imaging Assay for Characterizing Ca2+ Flux with R-GECO Biosensor

Expressed biosensors and automated imaging present a powerful system for conducting calcium mobilization studies. This application note describes the use of the R-GECO Ca2+ biosensor along with the Agilent BioTek Lionheart FX and Cytation imaging systems to detect and quantify receptor activation and resulting calcium flux within HEK 293 cell culture.

Read app note