p53 (Acetyl-Lys317)Transcription Factor Activity Assay

This DNA-Binding Transcription Factor Activity Assay from Assay Genie quantitatively measures p53's ability to bind its consensus DNA response element when the protein is acetylated at lysine 317 (Ac-Lys317), a C-terminal regulatory modification linked to transcriptional activation of cell-cycle arrest and apoptotic target genes. A biotinylated double-stranded oligonucleotide carrying the p53 response element is immobilized on streptavidin-coated wells; activated p53 from nuclear extracts binds the immobilized DNA and is detected with a rabbit anti-Ac-Lys317 primary antibody and an HRP-conjugated secondary, producing an absorbance-based readout. The kit is validated for human, mouse, and rat nuclear and cell lysates, with a recommended loading of approximately 10-20 uL per well of clarified nuclear extract at 1-2 mg/mL total protein. Store all components at -20 degrees C, protect the antibody vials from light, and limit freeze-thaw cycles of reconstituted reagents to preserve Ac-Lys317 signal; the microplate format supports up to 96 independent binding reactions per kit.