| Assay Method Information | |
| | Biologic and Pharmacologic Assay |
| Description: | The media was removed and replaced by 50 μL of 1× lysis buffer from the AlphaLISA kit (with 1× of cOmplete™ protease inhibitor). The plates were sealed, and cells were allowed to lyse for 10 min in the shaker before storing the plate at −20° C. until used. The AlphaLISA assay with processed cell lysates were performed as described in the protocol. Briefly, 10 μL of the undiluted sample were loaded to 384 well Optiplates and 5 μL of the acceptor mix (as described in the AlphaLISA kit) was added to each well. The plates were sealed and briefly vortexed followed by the incubation at RT for 3 hours. In the dark, 5 μL of donor mix was added to each well. All plates were sealed and stored at RT overnight. The plates were read in EnVision reader using standard AlphaLISA settings.Compound activity was analyzed using CBIS data analysis suite (ChemInnovation, CA). |
| Affinity data for this assay | |
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