Tumor suppressor protein p53 is one of the most important regulators of cell proliferation, differentiation and programmed cell death, triggering growth arrest and/or apoptosis in response to different cellular stress signals. The sequence-specific DNA binding function of p53 protein can be activated by several different stimuli modulating the C-terminal domain of this protein. The most likely physiological mechanism of p53 sequence-specific DNA binding activity is its post-translational modification, mainly phosphorylation of specific sites. Here we used non-radioactive electrophoretic mobility shift assay (EMSA) to show that C-terminal phosphorylation of p53 protein by cdk2/cyclin A at serine 315 and by PKC at serines 371, 376 and 378 can efficiently stimulate p53 binding to DNA in vitro as well as binding of C-terminal monoclonal antibody Bp53-10 recognizing the residue 371-380.