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p53 expression in normal paraffin-embedded tissue using different antibodies and antigen retrieval buffer systems.
G Pillai1, H Roberts, K Gatter
1Nuffield Department of Clinical Laboratory Sciences, John Radcliffe Hospital, Oxford, UK. gaetha.pillai@ndcls.ox.ac.uk
Histopathology
|December 21, 2002
Summary
The DO7 antibody successfully detected wild-type p53 in normal paraffin-embedded tissues, showing nuclear and cytoplasmic staining. The 1801 antibody did not yield detectable p53 in these tissues.
Area of Science:
- Immunohistochemistry
- Molecular Biology
- Pathology
Background:
- Wild-type p53 protein is crucial in cellular processes.
- Detecting wild-type p53 in normal tissues is important for research.
- Standardized methods are needed for reliable p53 detection.
Purpose of the Study:
- To evaluate the efficacy of DO7 and 1801 antibodies for detecting wild-type p53.
- To assess the impact of different antigen retrieval buffers on p53 detection.
- To demonstrate wild-type p53 presence in normal paraffin-embedded tissues.
Main Methods:
- Utilized formalin-fixed paraffin-embedded normal tissue samples.
- Employed two antibodies: DO7 and 1801.
- Tested three antigen retrieval buffer systems (pH 9.8-9.9, pH 6.0, pH 8.0) with microwave treatment.
Main Results:
- DO7 antibody showed significant nuclear staining in various normal tissues (intestine, lymphoid, neurons, prostate, cardiac, bronchial).
- Cytoplasmic staining was observed with DO7 in prostate, pancreas, and kidney.
- The 1801 antibody failed to detect p53 in any of the tested normal tissues.
Conclusions:
- Wild-type p53 is present in normal paraffin-embedded tissues.
- DO7 antibody is effective for detecting wild-type p53 in these tissues.
- DO7 is recommended over the 1801 antibody for p53 detection in normal tissues.