- Epitope and Antibody Analysis
- Infectious Disease Research
- Autoimmune Research
- Cancer Research
- Epitope Mapping
- Conformational Epitope Mapping
- Antibody Cross-Reactivity Analysis
- Epitope Substitution Scan
- Antigen and Epitope Discovery
- Serum Biomarker Discovery
- Immune Monitoring
- Kinase Substrate Optimization
- Peptide Target Binder Development
Peptide microarrays are a valuable tool for kinase substrate optimization. The peptide array covers a single substitution scan of the phosphorylation site and is incubated with the active tyrosine kinase or serine/threonine kinase in the presence of ATP. Phosphorylated tyrosines, serines or threonines will be selectively stained with a fluorescently labeled tag. Peptide variants that are less or stronger phosphorylated than the original substrate can be identified.
Phosphorylation of two peptide substrate families by c-Src. The sequences of the strongest peptide variants were listed as top hits. Peptide substrates were further classified according to their spot intensities in background (-), weak (1,000-3,000 FU; +), medium (3,000 – 10,000 FU; ++), strong (10,000 – 20,000 FU; +++) and very strong (> 20,000 FU; ++++) signals; amino acid exchanges compared to the starting substrate were shown in green.
- Array content: Successive replacement of the amino acid positions -2, -1, +1, +2 next to the phosphorylation site of the starting peptide by all 19 other L-amino acids
- Control peptides: HA peptides in the array frame and polyEY and Poly EAY in the center of the array
Input Data & Sample Material
Peptide sequence of kinase substrate
100 µg purified and active kinase
Kinase substrate optimization is provided along with our comprehensive PEPperMAP® Service to ensure optimal results.