Protein Kinase Assay Kit, Universal

The Sigma Universal Protein Kinase Assay Kit is designed for flexibility in protein kinase activity assays. This kit allows the user to choose his or her own protein kinase, peptide substrate and kinase reaction buffer. The kit may be used to assay activity of a protein kinase sample, or potency of a test substance or drug candidate in activating or inhibiting a protein kinase. Furthermore, the high efficiency and consistency of the ultrafiltration separation process across a large concentration range of the biotinylated peptide substrate makes it ideal for quantitative kinetic studies such as measuring the enzymatic turnover rate and Michaelis-Mentin constant (Km).

Uses affinity binding and ultrafiltration separation to analyze a sample simply, safely and consistently.
In this simple and efficient protein kinase assay, a protein kinase sample is combined with a biotinylated peptide substrate and ³²P-ATP. The components are allowed to react, then free avidin is added which binds to the biotinylated ³²P-peptide product. The sample is then subjected to a centrifugal ultrafiltration process using a membrane that retains the product-avidin complex and passes the free ³²P-ATP. Separation by such affinity ultrafiltration process is easy, gentle and efficient because the separation binding occurs in a solution that is relatively free of nonspecific background. In addition, the ultrafiltration separation is performed in a contained and compact system that eases the handling of the radioactive sample and reduces the radioactive waste. The sample reservoir of the centrifugal ultrafiltration unit containing the ³²P-peptide product as retentate can be directly placed into a liquid scintillation vial for counting.

sufficient for 50 reactions