Detecting p53 isoforms at protein level

Methods Mol Biol. 2013:962:15-29. doi: 10.1007/978-1-62703-236-0_2.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western / methods*
  • Cell Line
  • Drosophila
  • Gene Expression
  • Genes, Reporter
  • Humans
  • Immunoprecipitation / methods*
  • Luciferases / genetics
  • Luciferases / metabolism
  • Models, Animal
  • Protein Isoforms / analysis
  • Protein Isoforms / genetics
  • Transcriptional Activation
  • Tumor Suppressor Protein p53 / analysis*
  • Tumor Suppressor Protein p53 / genetics*
  • Zebrafish

Substances

  • Protein Isoforms
  • Tumor Suppressor Protein p53
  • Luciferases