Small-molecule Screening

These screens use purified proteins and substrates in buffered solutions to produce an optical (fluorescent, luminescent, etc.) readout that monitors enzymatic or binding activity. In these types of screens, small compound inhibitors are ranked by the SIGNALS, referring to the function of targeted protein.

Examples:

• FRET substrates for proteases

• Fluorophore-tagged glycosides for glycosidase inhibitors

• Fluorophore-tagged ligands for protein binding inhibitors

• Luciferase fragment-tagged proteins to measure the dimerization 

• Luciferase fragment-tagged proteins to measure the protein-protein interaction 

• HDAC-Glo for HDACs

• NADP-Glo for NADP-utilizing enzymes

These screens can be macromolecular or small-molecule screens that utilize cells plated in 96 or 384 or 1536 well plates to produce visual phenotypic or signal changes in the cells. The measurement can be a signal- or image-based readouts.

Examples:

• Cytotoxicity evaluation using CellTiterGlo

• Promoter-driven expression of luciferase (firefly, NanoLuc, Lucia) 

• Promoter-driven expression of fluorescent proteins 

• Luciferase fragment-tagged proteins to measure dimerization or protein binding

• Fluorescent protein-tagged proteins to measure expression of proteins

• Fluorescent protein-tagged proteins to monitor localization