Development of a sensitive multi-well colorimetric assay for active NFκB

Patricia Renard, Isabelle Ernest, Andrée Houbion, Muriel Art, Hervé Le Calvez, Martine Raes, José Remacle

Research output: Contribution to journalArticlepeer-review


The transcription factor nuclear factor kappaB (NFkappaB) is a key factor in the immune response triggered by a wide variety of molecules such as inflammatory cytokines, or some bacterial and viral products. This transcription factor represents a new target for the development of anti-inflammatory molecules, but this type of research is currently hampered by the lack of a convenient and rapid screening assay for NFkappaB activation. Indeed, NFkappaB DNA-binding capacity is traditionally estimated by radioactive gel shift assay. Here we propose a new DNA-binding assay based on the use of multi-well plates coated with a cold oligonucleotide containing the consensus binding site for NFkappaB. The presence of the DNA-bound transcription factor is then detected by anti-NFkappaB antibodies and revealed by colorimetry. This assay is easy to use, non-radioactive, highly reproducible, specific for NFkappaB, more sensitive than regular radioactive gel shift and very convenient for high throughput screening.

Original languageEnglish
Pages (from-to)1-5
Number of pages5
JournalNucleic Acids Research
Issue number4
Publication statusPublished - 15 Feb 2001


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