The pandemic coronavirus disease 2019 () is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As a first step of the viral replication strategy, the virus attaches to the host cell surface before entering the cell. The Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 may offer some protection against the viral infection.The SARS-CoV-2 Spike S1:ACE2 TR-FRET Assay is designed to measure the inhibition of the binding between SARS-CoV-2 Spike S1 and human ACE2 in a homogeneous 96 or 384 reaction format. This TR-FRET-based assay requires no time-consuming washing steps, making it especially suitable for high throughput screening applications. The assay procedure is straightforward and simple; the test inhibitor compound is incubated with biotinylated Spike S1, Eu-labeled ACE2, dye-labeled acceptor and an inhibitor for one hour. Then the TR-FRET signal is measured using a fluorescence reader.