Protocols and tools
The method we recommend to assess TP53 mutation status is to perform gene sequencing. Immuno-histochemical analysis is not a good method for assessing TP53 mutation status, as it cannot distinguish between the various types of mutations, does not detect truncating mutations (false-negative results), and gives false-positive results. Although most mutations cluster within exons 5-8, it is recommended to screen at least exons 4–10, including the splice junctions, so that less than 1% of mutations may be missed.
The standard sequencing procedure used at IARC for human TP53 gene is direct sequencing.
Protocols can be downloaded below:
- Amino-acid letter codes and properties
- Genetic code
- Human Splicing Finder
- Align-AGVGD (prediction of functional impact of missense mutations)
- SIFT (prediction of functional impact of missense mutations)
- Scorecons (for scoring residue conservation)
- Mutalyzer, a package to support checks of sequence variant nomenclature according to the guidelines of the Human Genome Variation Society
- Monoclonal antibodies targeting p53