Comparison of anti-p53 antibodies in immunoblotting.
Some of the most important tools to study p53 protein are various anti-p53 antibodies and immunological methods based on antibody-antigen reactions. Critical comments on the specificity and sensitivity of anti-p53 antibodies have been published. Four monoclonal and two polyclonal anti-p53 antibodies, four of them from two different sources, were compared for their ability to detect in immunoblotting the benzo(a)pyrene-induced p53 from C57BL/6 mouse skin and MCF-7 human breast carcinoma cells. Multiple extra bands were seen with most antibodies. A theoretical comparison of the equivalent epitopes of p53 homologues with the known epitopes of p53 antibodies indicated that the extra bands seen with most antibodies are not due to cross-reactivity with these homologues. A careful adjustment of antibody dilutions is needed for each application utilizing commercial p53 antibodies, regardless of the recommendations of the supplier.
Turpeinen
Miia
M
Department of Pharmacology and Toxicology, P.O. Box 5000, FIN-90014 University of Oulu, Oulu, Finland.
Serpi
Raisa
R
Rahkolin
Mika
M
Vähäkangas
Kirsi
K
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology; D000939:Epitopes / Q000737:chemistry; D005260:Female; D006801:Humans; D015151:Immunoblotting; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000276:immunology*
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000939:Epitopes; D016159:Tumor Suppressor Protein p53