Nonradioactive Kinase Assay Kits

Safety concerns and economic considerations have fueled a growing trend in the biomedical sciences: to shun the use of radioactivity when practical. Nonradioactive options for numerous applications have become widely available, including a number of nonradioactive kinase assay kits. Assays from different manufacturers employ a wide range of strategies. Most of these kits utilize antibodies, but two nonimmunochemical approaches use fluorescently tagged substrates and employ different means of separating phosphorylated and nonphosphorylated substrate fractions.

Kits using immunochemical reagents employ either "general use" antiphosphotyrosine preparations or phosphospecific antibodies that bind target antigens phosphorylated at specific serine (Ser), threonine (Thr) and/or tyrosine (Tyr) residues. Substrates may be random polymers, peptide substrates based on the phosphorylation sites of different proteins, phosphorylatable domains, or full-length substrate proteins. Some kits can be used for the assay of a broad group of enzymes within a given kinase family; others are tailor-made for...

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