The p53 tumour suppressor protein is one of the most important topics in cancer research. Its function is associated with the ability to bind DNA in a sequence-specific manner and to operate as a transcription factor. In the present study, we have developed a rapid and reliable method for analysing sequence-specific binding of p53 protein to DNA using a modified enzyme-linked immunosorbent assay (ELISA). In this p53/DNA-ELISA, we use streptavidin-coated microplates to capture biotinylated oligonucleotides containing p53 consensus sequences (p53CON). This newly developed nonradioactive assay allows the detection of p53/DNA complexes using different monoclonal antibodies recognising p53 and has comparable or higher sensitivity to more complicated radioactive methods. Using this method, we can detect binding of endogenous p53 to p53CON and activation of p53 protein for sequence-specific DNA binding. Variations of the basic protocol have also been developed to perform competition experiments and to study p53 binding to natural binding sequences. This modified DNA-ELISA is applicable for screening p53 binding properties from various sources in a short time.

• MeSH
• DNA metabolismus   MeSH
• ELISA metody   MeSH
• kompetitivní vazba MeSH
• konsenzuální sekvence MeSH
• lidé MeSH
• monoklonální protilátky MeSH
• nádorové buňky kultivované MeSH
• nádorový supresorový protein p53 genetika   imunologie   metabolismus   MeSH
• rekombinantní proteiny genetika   metabolismus   MeSH
• sekvence aminokyselin MeSH
• superhelikální DNA metabolismus   MeSH
• vazba proteinů MeSH
• vazebná místa genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA MeSH
• monoklonální protilátky MeSH
• nádorový supresorový protein p53 MeSH
• rekombinantní proteiny MeSH
• superhelikální DNA MeSH