We use FLIPR in a range of assay formats, readouts and target types
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.
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).
Human neutrophils were isolated from donor blood, incubated with calcium flux dye, dispensed into 384 well plates and treated with increasing concentrations of FMLF ligand.
A. Thalium flux assay used to measure potassium flux into cells. Thallium replaces potassium in the assay and once inside the cell, binds to a dye that becomes fluorescent.
B. FLIPR responses from cells expressing a potassium channel following stimulation to open the channel (A) or compounds that inhibit channel opening (B) or that enhance channel opening (C).
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.