| Assay Method Information | |
| | Surface Plasmon Resonance (SPR) |
| Description: | The surface plasmon resonance (SPR) experiments were conducted on a Biacore3000 (GE Healthcare). Myc-tagged cereblon was immobilized on a carboxymethylated dextran surface (CM5) amine coupled to anti-Myc antibody to recognize the Myc tag. His-tagged cereblon protein was immobilized on a carboxymethylated dextran surface with nitriloacetic acid (NTA), taking advantage of NTA/Ni2+ chelation. The prepared surface was allowed to equilibrate over three hours in running buffer (10 mM HEPES buffer @pH 7.4, 150 mM NaCl, 0.005% P20,2% DMSO). All compounds were prepared in 100% DMSO stock plates with a top concentration of 5 mM in a 3× serial dilution. Compounds were transferred from the stock plate to the assay plate and diluted into running buffer containing no DMSO. All compounds were run as a six-concentration series with a final assay top concentration of 100 μM.Data analysis was performed in Scrubber 2 (BioLogic software, Campbell, Australia). |
| Affinity data for this assay | |
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