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

Part# TB132
Printed in USA. Revised 5/09.
Instructions for Use of Products V5330 & V5340.

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 utilize brightly colored, fluorescent peptide substrates that are highly specific for the kinases in question. The hot pink color is imparted by the addition of a dye molecule to the PepTag^® Peptide substrate. Phosphorylation by PKC or PKA of their specific substrate alters the peptide’s net charge from +1 to –1. This change in the net charge of the substrate 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.