| Assay Method Information | |
| | ALK Kinase Inhibition Test |
| Description: | The specific experimental process was as follows: a test compound was formulated into a 10 mM mother solution with DMSO, stored at −20° C. after sub-packaging; the mother solution was thawed before use, diluted to 1 mM with DMSO, and then diluted with 1× kinase buffer (containing 50 mM 4-hydroxyethyl piperazine ethanesulfonic acid pH 7.0, 0.02% NaN3, 0.01% BSA, 0.1 mM ortho-vanadate, 5 mM MgCl2, 1 mM DTT, 37.5 nM supplementary enzyme buffer) by 40 times. 4 μL of the diluted solution of the test compound and 2 μL of 0.4 ng/well ALK kinase solution (Thermo Scientific) were added into reaction wells in turn, incubated for 10 min at room temperature, then 4 μL of 0.7 μM biotin-labeled tyrosine kinase substrate and 5 μM ATP solution were added to start the kinase reaction. After the reaction, 5 μL of 43.75 nM streptavidin-labeled XL665 was added, the materials were evenly mixed and then 5 μL of 0.04 nM europium-labeled tyrosine kinase antibody detection solution was imeddiately added. After reacting at room temperature for 1 hour, a fluorescence signal was detected using a SpectraMax i3× instrument (excited at 320 nm, emitted at 665 nm, 615 nm). |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |