Thomas Martinez

Clyde Calcium Assays

The B-lymphocyte Ca2+ leak assay is a simple, inexpensive, and reproducible assay that can be used for serial patient follow-up. More studies are needed to determine its accuracy and reproducibility, as well as its correlation with the progression of LVAD. Systemic neurohormonal activation results in accelerated disease progression. To determine its accuracy and reproducibility, large studies are needed. The results of these studies should be correlated with systemic neurohormonal activation, which is a key factor in LVAD progression.

Fluo-4 Direct(tm) calcium assay

The Fluo-4 Direct(tm) Calcium Assay measures intracellular calcium response in HEK 293 cells. The kit uses a calcium-sensitive dye called Fluo-4 DirectTM. The cells were seeded into 384-well microplates with 15,000 cells/well in 25 ul assay medium. Then, cells were infected with SFTSV at an MOI of five or twenty, or mock treated, and intracellular Ca2+ concentrations were determined with the fluorescence excitation and emission wavelengths of the dye.

The fluorescence signal was measured using a direct optical bottom reading method with a BMG LABTECH microplate reader. The calcium response was measured after a high resolution cell layer scan with the EX/EM 485/520 optical module. The 10x10 scan matrix used in this experiment helped correlate the calcium signal with the cell layer. The results of the assay were consistent with those of the controls.

The Fluo-4 Direct calcium flux assay was developed by Life Technologies. This commercially available calcium flux measurement kit was developed to monitor calcium flux in cells. Life Technologies patented Fluo-4 Direct Calcium Assay for its reagent. The assay is sensitive enough to measure calcium flux in a variety of cell types. For a more accurate measurement, the Fluo-4 Direct Calcium Assay should be used in combination with a cell culture assay.

CellOPTIQ(r) HCA

The CellOPTIQ(r) High Content Assay (HCA) is a new cardiac cell function assay developed by Clyde Biosciences. It enables the assessment of three variables in the same cells: calcium, phosphorus and myocardial contractility. These tests are trusted by world-leading biopharmaceutical companies and academic centers to identify cardiotoxicity in small molecule drug candidates.