PPARα transcription factor assay offers a non-radioactive method for detecting PPARα binding activity in nuclear extracts. A 96-well enzyme-linked immunosorbent assay (ELISA) replaces the radioactive electrophoretic mobility shift assay (EMSA). A specific double-stranded DNA (dsDNA) sequence containing the peroxisome proliferator response element (PPRE) is immobilized onto the wells of a 96-well plate. PPARs contained in a nuclear extract bind to the PPRE and PPARα is selectively detected using a specific primary antibody. A secondary antibody conjugated to HRP is added to provide colorimetric readout at 450 nm.