| Assay Method Information | |
| | Inhibition Assay |
| Description: | VEGFR2: Z'-LYTEC Kinase Assay Kit—Tyrosine 1 Peptide (PV3190, Invitrogen) was used to assay enzyme activity. 5 μl of recombinant human VEGFR2 enzyme (PV3660, Invitrogen) and VEGFR2 substrate polypeptide (in the reaction system, the final concentration of enzyme was 0.14 ng/μL, and the final concentration of substrate was 2 μM) formulated with reaction buffer (50 mM HEPES pH7.5, 10 mM MgCl2, 1 mM EGTA, 0.05% BRIJ-35), 2.5 μl of a 2-fold gradient dilution of the compounds dissolved in 5% DMSO, and 2.5 μL of ATP solution (the final concentration of ATP was 50 μM) were added successively to a 384-well plate (4513, Corning). After reaction at 25° C. for 2 hours, 5 μL of detection reagent was added to each well. After the plate was placed at 25° C. for 1 hour, the fluorescence signal values at emission wavelengths of 445 nm and 520 nm were measured with a NOVOstar (BMG) multi-function microplate reader. The IC50 values of the compounds for enzyme inhibition were calculated using Graphpad prism software based on each concentration of the compound and the corresponding signal value thereof. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |