| Assay Method Information | |
| | In Vitro Activity Test on Mutant TRKA (G595R) |
| Description: | Table 3: Experimental MaterialsThe recombinant human TRKA (G595R), TRKA (G667C) and TRKC (G623R) proteins were purchased from SignalChem. HTRF kinEASE TK kit was purchased from Cisbio Bioassays. Synergy Neo 2 of Biotek was used to read the plate. TRKA (G595R) Enzyme Reaction:0.12 ng/μL TRKA (G595R) kinase, 1 μM TK Substrate-biotin polypeptide substrate, 4.5 μM ATP, 1× enzymatic buffer, 5 mM MgCl2, and 1 mM DTT. The assay plate was White Proxiplate384-Plus plate (PerkinElmer), and the reaction was incubated at room temperature for 30 min, and the assay volume was 10 μL. Detection Steps:10 μL of detection reagent (containing 0.125 μM SA-XL665 and 5 μL 1×TK-Antibody) was added to the plate and incubated overnight at room temperature, and Synergy Neo 2 was used to read the plate. |
| Affinity data for this assay | |
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