Tyr'd and True: Immunochemical reagents and kits for studying tyrosine phosphorylation

Tyrosine Phosphorylantion State-Sensitive Antibodies Protein tyrosine kinases (PTKs) are a diverse lot, including both transmembrane and soluble cytoplasmic enzymes. Roughly 10 percent of a cell's proteins are subject to covalent modification via phosphorylation, but tyrosine (Tyr) residues are modified in only about one out of every 100 protein phosphorylation events. When Tyr-phosphorylation occurs, cells pay heed. PTKs play vital roles in a wide variety of cellular processes, including gro

Deborah Wilkinson

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If you are in the business of detecting, visualizing, or purifying Tyr-phosphorylated proteins, then antiphosphotyrosine antibodies [ anti-(P)Tyrs] will definitely be on your shopping list. Several types of anti-(P)Tyrs are commercially available. Polyclonal preparations are sold by various vendors, and a number of different monoclonal anti (P)Tyrs are also on the market. Most of the currently available anti-(P)Tyrs bind free (P)Tyr quite well--with affinities in the µM range or better. Cross-reaction with phosphoserine and phosphothreonine [(P)Ser and (P)Thr] is not generally a problem with these immunoreagents, but monoclonal anti- bodies generated in some of the early efforts can bind to nucleotides and (P)His. For a review of commercially available anti-(P)Tyr reagents and kits that contain them, read on.

Immunohistochemical staining with mAb 4G10. Image provided by Upstate Biotechnology.

In the 1980s, different research groups were trying out various antigens for ...

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