10.5281/zenodo.1308263
https://zenodo.org/records/1308263
oai:zenodo.org:1308263
Wong, Jong Fu
Jong Fu
Wong
0000-0001-6539-5865
Structural Genomics Consortium
Bullock, Alex
Alex
Bullock
0000-0001-6757-0436
Structural Genomics Consortium
Determining the IC50 of M4K1062 with ACVR1/ALK2-NanoLuc and different concentrations of Tracer-6908
Zenodo
2018
nanoBRET
ACVR1
ALK2
Target Engagement
Tracer-6908
M4K
2018-07-10
10.5281/zenodo.1308262
https://zenodo.org/communities/sgc-opennotebook
https://zenodo.org/communities/openlabnotebooks
Creative Commons Attribution 4.0 International
The EC50 of Tracer-6908 with ACVR1-c-nanoLuc and ideal conditions for the target engagement assay have been determined in the previous post. However, it is still necessary to verify that the IC50 values determined in the assay are closed enough approximation to the actual IC50 values of the compounds. If the IC50 values are strongly influenced by the concentration of Tracer-6908 used, multiple assays with progressively lower Tracer-6908 will need to be performed. The actual IC50 of the compounds can then be estimated by plotting the Tracer-6908 concentrations against the corresponding IC50 values and regressing to no Tracer-6908. To address this, multiple nanoBRET target engagement assays were setup with increasing concentrations of Tracer-6908 with the same serial dilution of M4K1962 (starting from the EC50 of 65nM).