Substrate Identification
Kinase of interest is incubated with biotinylated substrate peptides. Biotinylated peptides are captured on streptavidin plates. Cell Signaling Technology antibodies are used as primary detection reagents to detect phosphorylated peptides (secondary antibody sold separately).
In order to expedite the identification of peptide substrates for novel kinases, Cell Signaling Technology has developed two Kinase Substrate Screening Kits. Intended to be used with uncharacterized kinases, the Kinase Substrate Screening Kits enable drug discovery researchers to quickly find substrates and corresponding phosphorylation-specific antibody detection reagents. Antibody and matched peptide substrate can then be used in HTS kinase assays.
These kits contain matched pairs of phosphorylation-specific antibodies and biotinylated peptides, and have been used successfully in numerous HTS assay formats (heterogeneous/homogeneous TRF, colorimetric ELISA, AlphaScreen, etc.). Signal-to-Noise ratios have been optimized using Europium labeled secondary antibody conjugates (available from PerkinElmer Life Sciences). Substrate sequences are derived from physiologically relevant peptides.
