NFYA Transcription Factor Activity Assay

The NFYA Transcription Factor Activity Assay from Assay Genie is a DNA-binding ELISA that quantifies the DNA-binding activity of Nuclear Factor Y subunit alpha (NF-YA) in nuclear and cell lysates. The plate is pre-coated with a double-stranded oligonucleotide containing the canonical CCAAT-box consensus sequence recognized by the NF-Y trimer (NF-YA, NF-YB, NF-YC).

Activated NF-Y present in nuclear extracts binds the immobilized probe, and bound transcription factor is detected using a specific primary antibody in an ELISA-based format. This approach lets researchers measure endogenous transcription factor activation without the time and radioisotope requirements of gel-shift or EMSA assays.

Common research applications include studies of cell cycle regulation (NF-Y directly activates cyclin B, cdc2, and E2F target genes), MHC class II gene expression, cancer biology where NF-Y is frequently dysregulated, and stem cell pluripotency pathways.

Key specifications: reactive with human, mouse, and rat samples; compatible with nuclear and whole-cell lysates; each kit provides reagents sufficient for 96 assays. Nuclear extracts are typically prepared by hypotonic lysis followed by high-salt extraction, with 5-20 ug of nuclear protein recommended per well. Store kit components refrigerated at 4 degrees C; for long-term storage, freeze aliquots at -20 degrees C to preserve antibody activity.