Mouse monoclonal clone PY20 anti-Phosphotyrosine is specific for both native and denatured proteins containing phosphorylated tyrosine. Antibody binding is inhibited with phosphotyrosine and phenylphosphate, but not phosphoeserine or phosphothreonine.
Protein phosphorylation is a basic mechanism for the modification of protein function in eukaryotic cells. Tyrosine phosphorylation is a rare post-translational event in normal tissue, accounting for only 0.03% of phosphorylated amino acids. The level of phosphorylated tyrosine in many cellular proteins increases tenfold following various activation processes that are mediated through phosphotyrosine kinases. The importance of tyrosine phosphorylation has been established by the demonstration that it is an integral response in many different mitogenic receptor systems.
Reversible phosphorylation of proteins is an important post-translational modification that plays a regulatory role in the expression of most proteins in the cells. Reversible phosphorylation at multiple serine, tyrosine and threonine residues mediate numerous signalling pathways in both prokaryotic and Eukaryotic cells. Cellular proteins with phosphorylated tyrosine increase many fold by the activation of tyrosine kinases. Most mitogenic receptor systems such as EGF, PDGF, insulin receptors contain tyrosine kinase domains that undergo autophosphorylation when receptors bind to the respective ligands. Tyrosine-specific protein kinase activity has also been described in many retroviral oncogene proteins. Cells transformed by these oncogenes contain elevated levels of phosphotyrosine. Many of the oncogenes found in mammalian oncogenic viruses encode tyrosine protein kinases that reside in the cellular cytoplasm. Others encode transmembrane receptors whose tyrosine phosphotransferase activity is stimulated by the binding of ligand to the extracellular domain. Monoclonal Anti-Phosphotyrosine is specific for both native and denatured proteins containing phosphorylated tyrosine.
Immunogen
phosphotyrosine-protein conjugate
Application
Monoclonal anti-phosphotyrosine antibody produced in mouse has been used in the detection of tyrosine phosphorylation in proteins by 2D western blotting and western blotting.
Physical form
Solution in 20 mM phosphate buffered saline, pH 7.5, containing 50% glycerol and 3 mM sodium azide
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