title: Selective recognition of the activated form of transcription factor NF-kappa B by a monoclonal antibody.
abstract: Transcription factor NF-kappa B is a central regulator of inflammatory, immune and acute phase reactions. It rapidly initiates the transcription of a wide variety of target genes in response to various pathogenic stimuli. Because NF-kappa B is predominantly controlled at a posttranscriptional level through association with the inhibitory I kappa B subunits, its activation cannot be monitored directly at a cellular level by means of detecting new mRNA or protein synthesis. In this study, we describe a monoclonal antibody, designated alpha-p65MAb, that recognizes an epitope which includes the nuclear location signal (NLS) of p65, the DNA binding subunit mainly responsible for the strong gene-inductory potential of NF-kappa B. alpha-p65MAb recognized human and rodent p65 only when I kappa B alpha was not bound to p65. Thus, the IgG3 selectively stained the activated, nuclear form of NF-kappa B in cultured cells. Unlike I kappa B, the MAb and its Fab fragments did not inhibit the DNA binding activity of NF-kappa B in mobility shift assays. We show that alpha-p65MAb is suitable to study the activation state of NF-kappa B in cryosections of tissues.
authors.affiliation: Institute of Biochemistry, University of Freiburg, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Kaltschmidt
authors.forename: B
authors.initials: B
authors.lastname: Kaltschmidt
authors.forename: T
authors.initials: T
authors.lastname: Henkel
authors.forename: H
authors.initials: H
authors.lastname: Stockinger
authors.forename: P A
authors.initials: PA
authors.lastname: Baeuerle
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000276:immunology* / Q000378:metabolism; D011485:Protein Binding; D051996:Transcription Factor RelA
chemical_list: D000911:Antibodies, Monoclonal; D016328:NF-kappa B; D051996:Transcription Factor RelA