The Chroma-Glo™ Luciferase Assay System is designed to generate luminescence from luciferases emitting two different colors (red and green) within a single plate well (or from a single sample) after the addition of a single reagent. Quantitating multiple reporter gene outputs within a single sample well in a high throughput setting has traditionally been difficult. Different reporter proteins require different, often incompatible environments. Additionally, sample well size limits the number of reagent additions possible. Filtered measurement of the luminescence produced by Chroma-Luc™ luciferases permits red- and green-emitting luciferases to be measured independently. This homogeneous assay generates luminescence with signal half-lives generally >30 minutes, which enables the processing of multiple plates without multi-step sample handling. Two pieces of information can thus be efficiently and reproducibly gleaned from each well in a typical high-throughput screen.