PepTag® Assay for Non-Radioactive Detection of Protein Kinase C or cAMP-Dependent...

Literature # TB132

The PepTag® Non-Radioactive Protein Kinase Assays provide a rapid, sensitive and non-radioactive method to detect cAMP-dependent protein kinase (PKA) and protein kinase C (PKC). The PepTag® Assays use brightly colored, fluorescent peptide substrates that are highly specific for the kinases in question. The hot pink color of the PepTag® Peptide substrate is imparted by addition of a dye molecule. Phosphorylation by PKC or PKA alters the peptide substrate’s net charge from +1 to –1. This change in the net charge allows the phosphorylated and nonphosphorylated versions of the substrate to be rapidly separated on an agarose gel. The phosphorylated species migrates toward the positive electrode, while the nonphosphorylated substrate migrates toward the negative electrode. Using the PepTag® Assay, less than 10ng of kinase can be detected in under 2 hours. This allows rapid screening of large numbers of samples such as those produced from the assay of multiple column fractions.