Nucleic Acids Research, 2001, Vol. 29, No. 4 e21
© 2001 Oxford University Press
Development of a sensitive multi-well colorimetric assay for active NF
B
Laboratoire de Biologie et Biochimie Cellulaire, Facultés Universitaires Notre-Dame de la Paix, 61 rue de Bruxelles, B-5000 Namur, Belgium and 1Active Motif, 5431-C Avenida Encinas, Carlsbad, CA 92008, USA
The transcription factor nuclear factor 
B (NFB) 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 NFB activation. Indeed, NFB 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 NFB. The presence of the DNA-bound transcription factor is then detected by anti-NFB antibodies and revealed by colorimetry. This assay is easy to use, non-radioactive, highly reproducible, specific for NFB, more sensitive than regular radioactive gel shift and very convenient for high throughput screening.
* To whom correspondence should be addressed. Tel: +32 81 724128; Fax: +32 81 724135; Email: patsy.renard{at}fundp.ac.be
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  D. J. Klinke II, I. V. Ustyugova, K. M. Brundage, and J. B. Barnett Modulating Temporal Control of NF-{kappa}B Activation: Implications for Therapeutic and Assay Selection Biophys. J., June 1, 2008; 94(11): 4249 - 4259. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Gustin, E. Delaive, M. Dieu, D. Calay, and M. Raes Upregulation of Pentraxin-3 in Human Endothelial Cells After Lysophosphatidic Acid Exposure Arterioscler. Thromb. Vasc. Biol., March 1, 2008; 28(3): 491 - 497. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Hou, Z. Chen, M. Ji, N. He, and Z. Lu Real-time PCR Assay for Ultrasensitive Quantification of DNA-Binding Proteins Clin. Chem., April 1, 2007; 53(4): 581 - 586. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Urbinati, A. Bugatti, M. Giacca, D. Schlaepfer, M. Presta, and M. Rusnati {alpha}v{beta}3-integrin-dependent activation of focal adhesion kinase mediates NF-{kappa}B activation and motogenic activity by HIV-1 Tat in endothelial cells J. Cell Sci., September 1, 2005; 118(17): 3949 - 3958. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. M. Necela and J. A. Cidlowski A Single Amino Acid Change in the First Zinc Finger of the DNA Binding Domain of the Glucocorticoid Receptor Regulates Differential Promoter Selectivity J. Biol. Chem., September 17, 2004; 279(38): 39279 - 39288. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Rouet-Benzineb, T. Aparicio, S. Guilmeau, C. Pouzet, V. Descatoire, M. Buyse, and A. Bado Leptin Counteracts Sodium Butyrate-induced Apoptosis in Human Colon Cancer HT-29 Cells via NF-{kappa}B Signaling J. Biol. Chem., April 16, 2004; 279(16): 16495 - 16502. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Banan, L. J. Zhang, A. Farhadi, J. Z. Fields, M. Shaikh, and A. Keshavarzian PKC-{beta}1 isoform activation is required for EGF-induced NF-{kappa}B inactivation and I{kappa}B{alpha} stabilization and protection of F-actin assembly and barrier function in enterocyte monolayers Am J Physiol Cell Physiol, March 1, 2004; 286(3): C723 - C738. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Yang, X. Li, T. W. Prow, L. M. Reece, S. E. Bassett, B. A. Luxon, N. K. Herzog, J. Aronson, R. E. Shope, J. F. Leary, et al. Immunofluorescence assay and flow-cytometry selection of bead-bound aptamers Nucleic Acids Res., May 15, 2003; 31(10): e54 - e54. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. A. Zimmerman, C. H. Selzman, L. L. Reznikov, S. A. Miller, C. D. Raeburn, J. Emmick, X. Meng, and A. H. Harken Lack of TNF-alpha attenuates intimal hyperplasia after mouse carotid artery injury Am J Physiol Regulatory Integrative Comp Physiol, August 1, 2002; 283(2): R505 - R512. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. A. Zimmerman, C. H. Selzman, L. L. Reznikov, C. D. Raeburn, K. Barsness, R. C. McIntyre Jr., C. R. Hamiel, and A. H. Harken Interleukin-11 attenuates human vascular smooth muscle cell proliferation Am J Physiol Heart Circ Physiol, July 1, 2002; 283(1): H175 - H180. [Abstract] [Full Text] [PDF]
|
|