PepTag® Non-Radioactive Protein Kinase Assays

The PepTag® Non-Radioactive Protein Kinase Assay Systems provide a rapid, sensitive and non-radioactive method to detect either Protein Kinase C (PKC) or Protein Kinase A (PKA) activity. The PepTag® Assays use brightly colored, fluorescent peptide substrates that are highly specific for PKC (PepTag® C1 Peptide-PLSRTLSVAAK) and PKA (PepTag® A1 Peptide-LRRASLG). Phosphorylation of the peptide alters the 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 at neutral pH. Using fluorescent detection, ...

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The PepTag® Non-Radioactive Protein Kinase Assay Systems provide a rapid, sensitive and non-radioactive method to detect either Protein Kinase C (PKC) or Protein Kinase A (PKA) activity. The PepTag® Assays use brightly colored, fluorescent peptide substrates that are highly specific for PKC (PepTag® C1 Peptide-PLSRTLSVAAK) and PKA (PepTag® A1 Peptide-LRRASLG). Phosphorylation of the peptide alters the 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 at neutral pH. Using fluorescent detection, less than 2ng of purified kinase can be detected in less than 2 hours. The PepTag® Non-Radioactive Protein Kinase Assay Systems can detect kinase activity in partially purified samples as well as purified preparations of enzymes, making it a good choice for the rapid screening of column fractions or the screening of kinase activators and inhibitors. In addition to the assay components, each system includes purified kinase for use as a positive control.

Features - Benefits

• Non-Radioactive: The fluorescent tag on the peptide substrate facilitates quantitation of the phosphorylation reaction without the use of radioactivity.
• Low Background: Because the phosphorylation of the colored peptide supplied with the system is used to measure kinase activity, phosphorylation of other substrates occurring naturally in the sample does not add to the kinase activity measured.
• Convenient: Quantitation of the phosphorylated peptide can be accomplished using a densitometer, spectrophotometer, 96-well plate reader, or fluorometer.

Applications

For more information, see the Protocols & Applications Guide.

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