title: The 4G10, pY20 and p-TYR-100 antibody specificity: profiling by peptide microarrays.
abstract: The reversible phosphorylation of tyrosine residues is one of the most frequent post-translational modifications regulating enzymatic activities and protein-protein interactions in eukaryotic cells. Cells responding to internal or external regulatory inputs modify their phosphorylation status and diseased cells can often be diagnosed by observing alterations in their qualitative or quantitative phosphorylation profile. As a consequence the ability to describe the phosphorylation profile of a cell is central to many approaches aiming at the characterisation of signalling pathways. Anti-phosphotyrosine (pY) antibodies are widely used as experimental tools to monitor the phosphorylation status of a cell. By using peptide microarray technology we have characterised the substrate specificity of three widely used pY antibodies. We report that they are more sensitive to sequence context than is generally assumed and that their sequence preferences differ.
authors.affiliation: Department of Biology, University of Rome Tor Vergata, Via della ricerca scientifica, 00133 Rome, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: Tinti
authors.forename: Aurelio Pio
authors.initials: AP
authors.lastname: Nardozza
authors.forename: Emanuela
authors.initials: E
authors.lastname: Ferrari
authors.forename: Francesca
authors.initials: F
authors.lastname: Sacco
authors.forename: Salvatore
authors.initials: S
authors.lastname: Corallino
authors.forename: Luisa
authors.initials: L
authors.lastname: Castagnoli
authors.forename: Gianni
authors.initials: G
authors.lastname: Cesareni
mesh_terms: D000595:Amino Acid Sequence; D045163:Antibodies, Phospho-Specific / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D057809:HEK293 Cells; D006801:Humans; D008969:Molecular Sequence Data; D010748:Phosphopeptides / Q000737:chemistry / Q000276:immunology; D010766:Phosphorylation; D040081:Protein Array Analysis / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D012815:Signal Processing, Computer-Assisted
chemical_list: D045163:Antibodies, Phospho-Specific; D010748:Phosphopeptides