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NF-Y is a ubiquitously expressed heterodimeric transcription factor composed of three subunits (NF-YA, NF-YB and NF-YC). All three subunits are required for constitution of a functional binding complex to the CCAAT box. CCAAT boxes are found in the regulatory region of 30% of genes, including housekeeping genes as well as inducible and cell cycle-regulated genes. The role of a CCAAT box as a cis-acting promoter element depends on its ability to interact with NF-Y. Methods to measure overall levels and modifications of the NF-YA subunit are crucial to understanding transcriptional regulation of the numerous CCAAT box-containing genes. The TransAM NF-YA Kit contains an antibody specific for the active form of the NF-YA subunit when it is bound to the target DNA.
Applications
NF-YA regulated genes are involved in a variety of cellular pathways that are currently being studied for new drug targets. Applications include:
- Drug screening and/or potency studies
- NF-YA transcriptional activity regulation
- Protein structure/function studies of NF-YA and its mutated variantss
Figure 1: Monitoring NF-YA activation with the TransAM NF-YA Kit.
Different amounts of nuclear extracts from Jurkat cells were tested for NF-YA activity by using the TransAM NF-YA Kit. Note that incubation with wild-type NF-Y competitor oligonucleotides reduces NF-YA binding by over 80%, while incubation with mutated NF-Y competitor oligos had no effect on NF-YA binding to DNA.
