title: Detecting p53 isoforms at protein level.
abstract: The human p53 protein isoforms are expressed in several cell lines and modulate p53 tumor suppressor -activity, mainly through modulation of gene expression (1-4). Thus, identifying the pattern of p53 isoforms expression in cell lines is a key step for future studies of the p53 network (5). At the moment, the detection of p53 protein isoforms is based on the use of a panel of antibodies allowing their identification by comparing their molecular weights and their detection pattern by different antibodies (6). Here, classical protocols supplemented with technical know-how are described to detect p53 protein isoforms at protein level by Western blotting and immunoprecipitation. Furthermore, a simple method to study the impact of p53 protein isoforms on p53 transcriptional activity through luciferase reporter gene assays is provided.
authors.affiliation: CR-UK Cell Transformation Research Group, Centre for Oncology and Molecular Medicine, Ninewells Hospital, University of Dundee, Dundee, UK.
authors.forename: Virginie
authors.initials: V
authors.lastname: Marcel
authors.forename: Marie P
authors.initials: MP
authors.lastname: Khoury
authors.forename: Kenneth
authors.initials: K
authors.lastname: Fernandes
authors.forename: Alexandra
authors.initials: A
authors.lastname: Diot
authors.forename: David P
authors.initials: DP
authors.lastname: Lane
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Bourdon
mesh_terms: D000818:Animals; D015153:Blotting, Western / Q000379:methods*; D002460:Cell Line; D004330:Drosophila; D015870:Gene Expression; D017930:Genes, Reporter; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D023421:Models, Animal; D020033:Protein Isoforms / Q000032:analysis / Q000235:genetics; D015533:Transcriptional Activation; D016159:Tumor Suppressor Protein p53 / Q000032:analysis* / Q000235:genetics*; D015027:Zebrafish
chemical_list: D020033:Protein Isoforms; D016159:Tumor Suppressor Protein p53; D008156:Luciferases