7.7 p53

Experimental Procedures

1,The p53-mediated cellular signal transduction pathway plays a crucial role in regulating the normal life activities of cells.
The p53-mediated cellular signal transduction pathway plays a vital role in regulating the normal life activities of cells, and the number of genes regulated by p53 has exceeded 160. p53 is the gene with the highest correlation with human tumors discovered so far. It exerts a tumor-suppressive effect in many types of tumors, inducing growth arrest or apoptosis depending on physiological conditions and cell types. In addition, as a transactivator, p53 is involved in the regulation of the cell cycle.

2,During the Western Blotting (WB) process, we must pay close attention to the expression level of the sample itself.
During the WB process, we must pay close attention to the expression level of the sample itself. For example, cells such as Hela, 293T, and Jurkat have no basal expression of p53 or express it at an extremely low level, making them suitable for transfection experiments. We have listed the expression status of p53 in some cell lines, as shown in the table below:
In case of no bands appearing in WB, it is essential to select appropriate cells from reliable sources for the experiment, and attention should be paid to the fresh preparation of samples. p53 is a nuclear protein; during lysis, ultrasonic treatment is strongly recommended, or a dedicated nuclear extraction kit can be used. In addition, p53 has multiple phosphorylation sites, acetylation modifications, and various isoforms. This is also the reason why multiple bands easily appear in some samples, and different isoforms are expressed in a tissue-dependent manner.