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™). 

KinaseFinder Assay Principle

List of Kinase Panels

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

Example data with 50 Ser/Thr kinases

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

Screening Details

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. 

Reference Publication

Corvin Walter et al. employed the Kinase Finder service to identify kinases that phosphorylate the TOM70, which represents the main import receptor for the carrier import pathway. Kinases DYRK1A and DYRK1B scored highest in phosphorylating TOM70 in the primary Kinase Finder screen (Fig 1a). The hits were confirmed by using increasing concentrations of substrate (Supp Fig. 1b). To confirm the DYRK1 specific phosphorylation site, a TOM70 mutant was used for screening which resulted in significantly less phosphorylation (Supp. Fig. 1g).