Development of a sensitive multi-well colorimetric assay for active NFkappaB.

@article{citeulike:1876925, abstract = {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.}, address = {Laboratoire de Biologie et Biochimie Cellulaire, Facult\'{e}s Universitaires Notre-Dame de la Paix, 61 rue de Bruxelles, B-5000 Namur, Belgium. patsy.renard@fundp.ac.be}, author = {Renard, P. and Ernest, I. and Houbion, A. and Art, M. and Le Calvez, H. and Raes, M. and Remacle, J. }, citeulike-article-id = {1876925}, issn = {1362-4962}, journal = {Nucleic Acids Res}, keywords = {nfkb}, month = {February}, number = {4}, posted-at = {2007-11-07 09:19:59}, priority = {2}, title = {Development of a sensitive multi-well colorimetric assay for active NFkappaB.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/11160941}, volume = {29}, year = {2001} }

TY - JOUR ID - citeulike:1876925 L3 - citeulike-article-id:1876925 TI - Development of a sensitive multi-well colorimetric assay for active NFkappaB. JF - Nucleic Acids Res VL - 29 IS - 4 AD - Laboratoire de Biologie et Biochimie Cellulaire, Facultés Universitaires Notre-Dame de la Paix, 61 rue de Bruxelles, B-5000 Namur, Belgium. patsy.renard@fundp.ac.be SN - 1362-4962 N2 - 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. KW - nfkb AU - Renard, P AU - Ernest, I AU - Houbion, A AU - Art, M AU - Le Calvez, H AU - Raes, M AU - Remacle, J PY - 2001/02/15/ UR - http://view.ncbi.nlm.nih.gov/pubmed/11160941 ER -