The KinaseFinder Screening Service
The KinaseFinder is a biochemical assay intended to identify kinases, which accept a given substrate for phosphorylation.
The KinaseFinder is a standardized service package for identifying protein kinases that can phosphorylate a peptide or protein of interest provided by our customer. Kinase hits, identified with the standard package, should subsequently be confirmed in a second experiment. The KinaseFinder is ideal for the elucidation of physiological pathways with the involvement of protein kinases.
- Potential substrates can be peptides or proteins
- Compare peptides with phospho-site mutations
- Option to follow up with an SDS-PAGE and autoradiogram to visualize the phosphorylated substrate
KinaseFinder Assay Principle
The target substrate is incubated with different kinases in every well of a multi-well plate and 33P-ATP that serves as a phosphate donor.
The kinase catalyzes the transfer of 33P when the protein or peptide is recognized as a substrate.
Substrates are either bound to scintillant-coated FlashPlates (33PanQinase™) or dropped on filter paper for readout (HotSpot™).
List of Kinase Panels
| Type of KinaseFinder | # of kinases | Assay format |
|---|---|---|
| Tyr protein kinases | 94 | ³³PanQinase™ |
| Ser/Thr protein kinases | 245 | ³³PanQinase™ |
| Ser/Thr & Tyr protein kinases | 339 | ³³PanQinase™ |
| Wild Type Kinase Panel | 376 | HotSpot™ |
| Custom panel | selected by customer | HotSpot™ |
Assay Details
Example data with 50 Ser/Thr kinases.
Activity of kinases with example substrate protein are shown (corrected for substrate background activity). Substrate protein was tested at one concentration. Yellow: Kinase with substrate protein; blue: Kinase without substrate protein (autophosphorylation). A ratio value between substrate phosphorylation and apparent autophosphorylation > 3 may be considered as significant. Performed with the 33PanQinase™ assay. cpm = counts per minute
Setups: Proteins and peptides are accepted as substrates.
Assay format: radiometric screening with 33P-ATP as phosphate donor
Turnaround time: 10 business days for standard projects. Expedited scheduling and data delivery can be arranged prior to commencement of the studies.
Screening location: Assays run with the 33PanQinase™ assay are performed at the German site. Assays run with the HotSpot™ assay are performed at the US site.
Related Resources
