Most kinase activity assays are binding assays used to determine a drug's effect on the phosphorylation of a protein substrate (for example, a peptide) by a specific kinase. A purified active kinase is used in an optimized buffer condition with the protein substrate. Phosphorylation of the substrate is detected using anti-phospho-antibodies labeled with fluorescent or radioisotopic probes.
This assay format is incompatible with a multiplex format for two reasons. First, it is difficult to optimize a buffer for a "universal" kinase assay. Second, you need a specific substrate for the kinase assay. To measure the activity of one specific kinase in experimentally treated cells, it cannot be labeled nonspecifically by other kinases.
The Bio-Plex Multiplex Phosphoprotein Assays are designed to provide a high-throughput means of simultaneously analyzing several phosphorylated kinases within the cell using a sandwich immunoassay format. The sandwich immunoassay is compatible with a multiplex format because it uses two antibody probes for the detection of the specific phosphorylation of a native kinase.
For more inforation, see Bulletin 5483, Bio-Plex Phosphoprotein Detection Performance.
Visit the Bio-Plex Multiplex Immunoassays page to view the entire range of available products.
To find assays for specific targets, use the Bio-Plex Assay Finder.