| Assay Method Information | |
| | Inhibitory Activity of Compounds on TRKA, TRKB, TRKC and ROS1 |
| Description: | The compound powder was dissolved in 100% DMSO (Sigma, Cat. D8418-1l) to prepare a 10 mM storage solution. The compounds had an initial test concentration of 100 nM, were 3-fold serially diluted to obtain 10 samples for multiple hole inspection. For TRKA, TRKB and TRKC kinase targets, Loxo-101 (Selleckchem, Cat. S7960) was used as positive reference compound; for ROS1, Staurosporine (Selleckchem, Cat. S1421) was used as positive reference compound. The gradient diluted compounds were mixed with TRKA/TRKB/TRKC/ROS1 kinase (Carna, Cat. 08-186/08-187/08-197/08-163) with final concentration of 2.5 nM/2.55 nM/2.5 nM/0.3 nM in a Optiplate-384F plate (PerkinElmer, Cat. 6007270), and incubated at room temperature for 10 minutes. After that, ATP was added with final concentration of 47.8 μM/71.2 μM/44.4 μM/26.7 μM, 3 μM Kinase Substrate22 (GL Biochem, Cat. 112393) was added. The reaction was carried out at room temperature for 30 min/40 min/20 min/20 min respectively. |
| Affinity data for this assay | |
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