p53 FUSION PROTEIN EXPRESSION IN PROKARYOTE AND PREPARATION OF MONOCLONAL ANTIBODY TO p53
Abstract
Objective: Conventional immunohistochemistry (IHC) is available to assess p53 mutations, and expensive imported anti-P53 monoclonal antibody has been used in China, it is necessary to study a new monoclonal antibody.
Methods: The p53 DNA fragment enconding N.terminal 180 amlao acide was obtained by PCR and was cloned into PGEX-2T plasmid expressing glutathione S.transferase (GST). The P53-GST fusion protein expressed by JM109 was used for immunizing BALB/C mice. We have raised one hybridoma strain secreting McAb to human p53 (named M126).
Results: The IHC analysis of 52 paraffin-embedded sections from human breast cancer with M126 and PAB1801 (Zymed Co.) has showed that the positive immunoreactions were 25 cases (48%) and 22 cases (42.3%) respectively. The staining of M126 was stronger and preferable to PAB1801.
Conclusion: M126 can be instead of PAB1801 for studying immunohistocbemica] analysis on p53 protein.
Methods: The p53 DNA fragment enconding N.terminal 180 amlao acide was obtained by PCR and was cloned into PGEX-2T plasmid expressing glutathione S.transferase (GST). The P53-GST fusion protein expressed by JM109 was used for immunizing BALB/C mice. We have raised one hybridoma strain secreting McAb to human p53 (named M126).
Results: The IHC analysis of 52 paraffin-embedded sections from human breast cancer with M126 and PAB1801 (Zymed Co.) has showed that the positive immunoreactions were 25 cases (48%) and 22 cases (42.3%) respectively. The staining of M126 was stronger and preferable to PAB1801.
Conclusion: M126 can be instead of PAB1801 for studying immunohistocbemica] analysis on p53 protein.