| Assay Method Information | |
| | AlphaLISA Assay |
| Description: | The means the half maximal inhibitory concentration of compound that inhibits CUL4A and DDB1-BPB binding. Our lab established and optimized 384 well-based AlphaLISA in vitro binding assay for testing small molecule inhibitory activity. Small molecule libraries were screened for compounds that disrupt CUL4A-DDB1-BPB interaction using AlphaScreen detection beads (PerkinElmer, #AL112C and #6765300) following the manufacturer's protocol (https://www.perkinelmer.com/PDFs/downloads/GDE-Alphatech.pdf, hereby incorporated by reference in its entirety for all purposes). The 384-well assay plates (Greiner Bio-one, #781075) were read in Synergy Neo2 Hybrid Multi-Mode Reader (BioTek). The IC50 was calculated by Prism 8 XY analysis nonlinear regression (curve fit-Log(inhibitor) vs response). |
| Affinity data for this assay | |
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