Steroidogenic factor 1 (SF-1) is a key regulator of endocrine function, especially steroidogenesis and reproduction. Unlike most nuclear receptors, SF-1 is constitutively activated and still remains an orphan receptor. To study its function, it is imperative to have reliable assays that can assess potential pharmacological modulators. Here we describe in detail three different cell-based assays that evaluate distinct aspects of SF-1 function: a cellular proliferation assay R-SAT® that monitors events far downstream of the receptor/ligand interaction, a transcriptional assay that focuses on the gene-modulating properties of SF-1, and an assay in adrenocortical cultures that constitutes a surrogate measure of SF-1 function in native tissues.