Conformational changes in p53 analysed using new antibodies to the core DNA binding domain of the protein. The use of these antibodies is subject to artefacts if they are not used carrefully. As shown below, PAb 1620 is very sensitive to the denaturation status of p53. Strong fixation procedure of cells can also lead to such loss. 1620 will never work on paraffin blocks and its reaction on frozen tissu should be carrefully controled.
All buffers contain protease inhibitors.
Washing of the immunoprecipitate was done in the corresponding lysis buffer. Mutations in p53 produce a common conformational effect that can be detected with a panel of monoclonal antibodies directed toward the central part of the p53 protein. Wild type versus mutant p53 accumulation in tissus. Several p53 mutants can have an unfolded structure in the tumor cells. This unfolding can lead to the unmasking of epitopes which can be detected on native p53.