P53 (tumor suppressor gene) or marker.
Can detect the mutation of p53 by IHC (monoclonal Ab) with out use PCR
Hi Abdullah,
The part of protein/antigen that binds to antibody is epitope. If any mutation causes conformational changes in epitope, only in such case, antibody will not bind to antigen (in your case p53). So,
1. Mutation in p53 anywhere else than antigen, should be undetected by antibody at all.
2. Mutation in epitope that result in conformational change avoiding antibody binding should be detected by absence of fluorescence.
3. Thus, Any other mutation than (#2) thus can not be detected by IHC.
Best,
Ankush
Hi Ankush,
Thank you so much for your interest.
So, do you have specific antibody use to detect the mutation of p53 gene.
What is the different between expression and mutation in IHC.
yes you can use this technique only without using PCR, the first one can identify how much p53 is over expressed as a protein the second one can help to deplicate the gene responsible for the mutation of P53.
Hi abdullah,
I always used antibody for WT_p53. But there are some groups that do work with mutation specific antibodies. For example: Article Immunohistochemical correlates of TP53 somatic mutations in cancer
Hope this helps!
The relation between TP53 mutation and TP53 accumulation is not 100%. Most missense TP53 mutation leads to an accumulation of TP53 protein in the nucleus of tumor cells. The most widely used antibody for staining tumors is DO1 or DO7 that binds efficiently to all TP53. Do not use any antibody specific for any mutation as they are not really efficient for tumor analysis. Keep in minds that any nonsense or frameshift mutations will not be detected via antibody staining. The best way is DNA sequencing. It is now very cheap.
Thierry
We have used DO7 in our study. Please see the publication (Nummela et al., Int J Cancer. 2015 Mar 1;136(5):E282-9. doi: 10.1002/ijc.29245) for classification of aberrant staining and representative figures.
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