Developed to perform quantitative optical screening for cell-based kinetic assays, Fluorescent Imaging Plate Reader (FLIPR) screening is used in a variety of assay formats, readouts and target types.
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We use FLIPR in a variety of assay formats, readouts and target types with a selection of examples listed below:
Antagonist and agonist screening recombinant cell-based assays
Cells expressing a target GPCR were loaded with calcium flux dye (Fluo-4) and treated with the GPCR ligand in the absence or presence of various compounds that bind to the GPCR and inhibit calcium response.
Dose response curve generated in a HEK-293 cell line expressing a 7-Transmembrane (7-TMR) or G-Protein-Coupled Receptor (GPCR) in a 384 well assay format
More recently, we have used both FLIPR Tetra and FLIPR Tetra (with the luminescent option) to successfully screen in 384 and 1536 well format using aqueorin cell lines purchased from PerkinElmer. We have experience of screening over a million compounds per assay in this format using ‘assay ready plates’ (first addition to the well is the compound).
Phenotypic screening of 7-TM receptos in human primary cells
Human neutrophils were isolated from donor blood, incubated with calcium flux dye, dispensed into 384 well plates and treated with increasing concentration of FMLF ligand.
Screening of compounds in Human Neutrophil FLIPR antagonist assay in 1536 well plates
Human neutrophils were isolated from the whole blood of donors by densite gradient centrifugation, loaded with Fluo-loading dye, dispensed into 1536 well plates incubated in the presence of compounds, and screened with agonist addition on-line. Responses from a selection of wells are shown in the image.
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