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Assay Method Information

Assay Name:  Counterscreen assay for S1P3 antagonists: Dose response cell-based high throughput screening assay to identify antagonists of the 5-Hydroxytryptamine Receptor Subtype 1E (5HT1E)
Description:  The CHO cell line was cultured in T-175 flasks at 37 degrees C and 95% relative humidity (RH). The growth media consisted of Dulbecco's Modified Eagle's Media containing 10% v/v heat inactivated dialyzed fetal bovine serum , 0.1 mM NEAA, 1 mM Sodium Pyruvate, 25 mM HEPES, 5 mM L-Glutamine, 1 mg/mL Geneticin, 5 micrograms /mL Blasticidin, 100 micrograms/ml Zeocin, and 1x penicillin-streptomycin-neomycin mix. Prior to the start of the assay, cells were suspended to a concentration of 0.4 million/mL in phenol red free Dulbecco's Modified Eagle's Media containing 10% v/v heat inactivated dialyzed fetal bovine serum, 0.1 mM NEAA, 1 mM Sodium Pyruvate, 25 mM HEPES, and 5 mM L-Glutamine. The assay was started by dispensing 10 ul of cell suspension to each well of a 384-well plate and incubating for 20 hours. Then, 50 nL of either test compound or DMSO control was added to the appropriate wells, in the presence of 1.1 ul of the selective 5HT1E agonist, BRL-54443 (3 nM final no
Affinity data for this assay
 

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Last update November 1, 2007
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