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FLIPR Screening
We have used FLIPR in a range of assay formats, readouts and target types including:
Antagonist and agonist screening in recombinant cell based assays
Working within 7-transmembrane receptor (7-TMR) receptor (also known as G-Protein Coupled Receptors or GPCR) biology for the last 15 years we have detailed knowledge and experience of developing assays and screening for 7-TMR activation and inhibition. We have used fluorescent methodologies with Fluo-3, Fluo-4, Fluo-4 no wash reagents and Fluo-5 in 96, 384 and 1536 well plates.
Working within 7-transmembrane receptor (7-TMR) receptor (also known as G-Protein Coupled Receptors or GPCR) biology for the last 15 years we have detailed knowledge and experience of developing assays and screening for 7-TMR activation and inhibition. We have used fluorescent methodologies with Fluo-3, Fluo-4, Fluo-4 no wash reagents and Fluo-5 in 96, 384 and 1536 well plates.
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 FLIPRTetra and FLIPRTetra (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).
More recently we have used both FLIPRTetra and FLIPRTetra (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 receptors in human primary cells
For some targets expressed endogenously in human primary cells (e.g. neutrophils) we have screened in 384 and 1536 well format using a fluorescent readout to prosecute the biology in its native environment. In this format we have screened up to 100,000 compounds per target.
For some targets expressed endogenously in human primary cells (e.g. neutrophils) we have screened in 384 and 1536 well format using a fluorescent readout to prosecute the biology in its native environment. In this format we have screened up to 100,000 compounds per target.
Screening of Compounds in Human Neutrophil FLIPR antagonist assay in 1536 well plates.
Human neutrophils were isolated from the whole blood of donors by density 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 selection of wells are shown in the image.
Human neutrophils were isolated from the whole blood of donors by density 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 selection of wells are shown in the image.
Measurement of protein-protein interaction
Using InteraX technology we have measured the degree of dimerisation between two proteins and used this approach to screen for compounds that inhibit the dimerisation process.
Using InteraX technology we have measured the degree of dimerisation between two proteins and used this approach to screen for compounds that inhibit the dimerisation process.
Inhibition of channel activation measured using flux dyes
Screening in 384 well format we have used dyes that traverse the channel to identify compounds that block the passage of ions through the channel.
Screening in 384 well format we have used dyes that traverse the channel to identify compounds that block the passage of ions through the channel.
These assays have been used as a first line screens to identify “hits” prior to more in-depth electrophysiological screening methods.