HNF4alpha/gamma Transcription Factor Activity Assay

This Transcription Factor Activity Assay from Assay Genie uses a biotinylated double-stranded oligonucleotide capture chemistry: biotin-labeled dsDNA probes carrying the HNF4 response element are immobilized on a streptavidin-coated plate via the high-affinity biotin-streptavidin interaction, after which nuclear or whole-cell lysate is added so that activated HNF4alpha and HNF4gamma dimers dock at the consensus site. Bound complexes are then recognized by isoform-specific primary antibodies and detected with an HRP-conjugated secondary antibody using TMB substrate, with the peroxidase reaction stopped using 2 N sulfuric acid before absorbance quantitation.

Each kit ships as twelve 8-well microstrips configured for 96 individual determinations and is validated for reactivity against human, mouse, and rat samples. Supplied reagents remain stable for 6 months when stored refrigerated at 4 degrees C, and the full workflow can be completed in roughly 4.5 hours from lysate loading to plate read. The HNF4alpha analyte maps to UniProt entry P41235 (gene symbol HNF4A, chromosomal locus 20q13.12, aliases TCF-14 and NR2A21), a nuclear receptor that binds DNA as a homodimer whose loss-of-function mutations are causally linked to Maturity-onset diabetes of the young type 1 (MODY1), supporting use of this kit in metabolic disease modeling, hepatocyte differentiation workflows, and dissection of hepatic gene regulatory networks. Research use only.