Assay Method Information

Assay Name:  Pharmacology Assay
Description:  Table 2: A study was conducted to determine the pharmacology of Compounds 1-18 in vitro in a plurality of biological matrices. Compounds 1-18 were prepared into proper concentrations of DMSO solution. The compound stock was then serially diluted into 10 concentrations by 3-fold dilution in 384 pp-plate. 5 nL of compound stock was transferred to a 384 plate. The compound stock was then diluted to 0.74 nM using a solution, which contained 0.79 mL of water, 0.2 mL of 5× Buffer, 2 μL of DTT, 6.66 μL of 7.5% BSA, and 5 μL of 10 mM ATP, to form an MKK7 mixture. The MKK7 mixture was then pre-warmed and 2.5 μL was added to each well designated for the “Test compound” and “High control” well, 2.5 μL solution 1 to each well designated for the “Low control” well. The well was incubated for 30 min and centrifuged at room temperature. Following centrifugation, 5 μL of ADD-Glo Reagent was added to each well and incubated for 60 min after centrifugation at room temperature. 10 μL of ADP-Glo Detection was then added to each well and incubated for 30 min after centrifugation in dark at room temperature.
Affinity data for this assay
 

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