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| | Dose Response Cell Based High Throughput Screening Assay for Agonists of the 5-Hydroxytryptamine Receptor Subtype 1E (5HT1E) |
| Description: | The CHO cell line was cultured in T-175 sq cm Corning flasks (part 431080) at 37 deg C and 95% RH. The HTS assay began by dispensing 10 microliters of cell suspension to each test well of a 384 well plate and incubating for 20 hours. Then 43 nL of either test compound or DMSO control (for negative control wells) was added. The selective 5HT1E agonist, BRL-54443 was also added (43 nL) to positive control wells to a final concentration of one micromolar, i.e. a concentration that resulted in 100% activity (EC100). Plates were then incubated at 37 deg C in 5% CO2 for 4 hrs. After the incubation, 2 microliter/well of the GeneBLAzer fluorescent substrate mixture (Invitrogen, LiveBLAzer, part K1096), prepared according to the manufacturer's protocol and containing 200 mM probenicid (Sigma, part P8761) was added to all wells. After 1 hour of incubation at room temperature, plate fluorescence was measured with the EnVision plate reader (PerkinElmer Lifesciences, Turku, Finland) at an exci |
| Affinity data for this assay | |
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