Assay Method Information

Assay Name:  KRAS Binding Assay
Description:  The ability of a test compound to bind to KRASG12D was measured by using an HTRF KRASG12D GTP Binding Kit (Cisbio, 63ADK000CB27PEG). 100×His-tagged Human KRASG12D stock solution was diluted to 1× by adding PPI Europium detection buffer and then transferring the diluted solution to a 384-well plate (5 μL/well, PerkinElmer, 6008289). 5 μL of compound solution was added to each well after serial dilution, followed by addition of 5 μL of GDP (50 nM) solution. After 30 minutes incubation at room temperature, 1×6His Eu cryptate antibody solution and GTP-Red tracer were added to the plate, then incubated for another 30 minutes. The HTRF signal was measured with a VICTOR Nivo multimode plate reader according to the manufacturer's instructions.
Affinity data for this assay
 

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