Kinase Panel Screening Service

Broad kinase selectivity screening is crucial for the identification of selective and potent kinase inhibitors. Determination and improvement of the selectivity of a compound by screening a large part of the kinome are of pivotal importance in the discovery process of novel kinase inhibitors e.g. to reduce the risk of adverse side effects or both, kinase inhibitors and compounds inhibiting non-kinase targets.

  • Reaction Biology offers the largest kinase panel for kinase profiling in the industry ensuring the most reliable results for your inhibitor selectivity screening covering most of the human kinome. 
  • Kinase profiling is performed on a monthly (³³PanQinase™) or bi-weekly (HotSpot™) schedule, which allows us to offer very competitive pricing.
  • Specialty panels are available for testing of compounds against a selection of kinase-specific mutants or our CDK panel. 
  • Kinase panel report from the US facility includes a free control compound's IC50 curve. Kinase panel reports from the German facility include the presentation of the kinase mapper for all wild type kinases profiled. 

Reaction Biology’s kinase assays were ranked highest in the Kinase Profiling Trends Survey by HTStec. Contact us today to discuss your project!

Visualize the Selectivity of your Compound

Broad kinase panel screening results can be visualized with our web-based kinase mapper tool. The selectivity profile of a compound can be easily compared to that of other compounds and patterns are readily visible. Try our tool here.

Kinase Mapper as tool for presenting kinase selectivity data

Selectivity Kinase Panels

Panels run at US facility
Panels run at US facility Panel size Panel
Wild Type Kinase Panel 374
Mutant Kinase Panel 267
Atypical Kinase Panel 24
Lipid Kinase Panel 17
Selectivity Kinase Panels
Panels run at German facility Panel size Panel
Wild Type Kinase Panel 335
Mutant Kinase Panel 94
Lipid Kinase Panel 14
IC50 Wild Type Kinase Panel 335

Speciality Kinase Panels

Panels run at US facility Panel size Data
CDK Profiler 29 Data sheet
ABL1 Mutant Profiler 13 Data sheet
ALK Mutant Profiler 22 Data sheet
BRAF Mutant Profiler 16 Data sheet
EGFR Mutant Profiler 44 Data sheet
FGFR2 Mutant Profiler 11 Data sheet
FLT3 Mutant Profiler 9 Data sheet
c-KIT Mutant Profiler 22 Data sheet
c-MET Mutant Profiler 27 Data sheet
RET Mutant Profiler 25 Data sheet
TIE Mutant Profiler 7 Data sheet
TRKA Mutant Profiler 15 Data sheet
TRKC Mutant Profiler 6 Data sheet
FGFR3 Mutant Profiler 6 Data sheet
FGFR4 Mutant Profiler 5 Data sheet
ERBB2 Mutant Profiler 8 Data sheet
Panels run at German facility Panel size Data
CDK Profiler 28 Data sheet
ABL1 Mutant Profiler 10 Data sheet
ALK Mutant Profiler 9 Data sheet
EGFR Mutant Profiler 10 Data sheet
KIT Mutant Profiler 9 Data sheet
MET Mutant Profiler 9 Data sheet
RET Mutant Profiler 11 Data sheet

Kinase Panel Screening Details

Assay Formats

HotSpot™ and ³³PanQinase™: two radiometric assay formats

Principle of radiometric kinase activity assay for kinase inhibitor screening.. Radioactive phosphate get's transferred to the kinase substrate. the now radioactive substrate get's quantified.

Reaction Biology offers two radiometric assay formats that are performed in our screening facilities located in Malvern, PA, USA, and Freiburg, Germany. Both assays are based on the transfer of ³³P-labelled phosphate from ATP to the kinase substrate. The methods, however, differ with respect to retention and detection of the phosphorylated substrates. Proteins and peptides in the HotSpot™ assay (performed in the US) are captured via spotting of the reaction mix on a filter membrane. The ³³PanQinase™ assay (performed in Germany) is performed with FlashPlate microtiter plates, which are coated with scintillant for detection.

Lipid kinases are screened with ADP-Glo (Promega) at both facilities. 

Reproducibility

Reproducibility IC50

The IC50 values of a BCR-ABL kinase inhibitor were determined for 320 wild type kinases in two independent experiments and compared. The high R2 value shows a high correlation of data. IC50 value determination was performed with 6 compound concentrations in singlicate with the ³³PanQinase™ assay.

 

IC50 determination

The control compound Staurosporine was tested over the course of 6 months in 12 independent experiments. IC50 determination was performed with 10 compound concentrations in singlicate with the HotSpot™ assay.  

Profiling at US facility

Screening formats: 2 data point projects with 1 concentration in duplicates and 5 to 10-doses response curves.

Assays: Protein kinases are screened via radiometric HotSpot™ assay. Lipid kinases are screened via ADP-Glo (Promega) assay. The assay protocol for HotSpot™ assay is available here.

ATP concentration: 1uM or 10 uM or apparent ATP-Km up to 100 uM

Controls: No inhibitor (DMSO vehicle) control. For every kinase, we include the screening of one kinase-specific control compound in 10-dose IC50 format.

Timelines: Panel screening is performed twice per month. Reports are sent 2 to 3 weeks after the project start.

Screening dates: Upcoming profiling dates are shown here

Report: The raw data, % enzyme activity and control compound IC50 values will be reported in Excel format. Assay conditions, target, and substrate information are available upon request. Requirements for this information should be noted prior to the commencement of the study.

Mapper: The results of the selectivity of compounds can be graphically presented with our mapper tool.   

Compound requirements: In brief, for kinase panel profiling 100 ul of a 10 mM DMSO stock or solid material is needed. Shipping information can be found in our FAQs.

Profiling at German facility

Screening formats: 2 data point projects are performed with 1 concentration in duplicates or 2 concentrations in singlicates. IC50 projects are performed with 6 concentrations in singlicate. 

Assays: Protein kinases are screen via radiometric ³³PanQinase™ assay. Lipid kinases are screened via ADP-Glo™ (Promega) assay. The assay protocol for the ³³PanQinase™ assay is available here

ATP concentrations: apparent ATP-Km

Controls: DMSO only as full enzymatic activity control.

Timelines: Panel screening is performed once per month or on-demand. Reports are sent 10 business days after the project start.

Screening dates: Upcoming profiling dates are shown here

Report: A detailed report will be provided including assay conditions, target, and substrate information. The raw data and % enzyme activity will be reported in Excel format including IC50 values and curve fitting, if applicable. Compound selectivity on wild type protein kinases will be graphically presented with the mapper tool.  

Compound requirements: For testing with 1 concentration in duplicate, 1700 µl of a 100x stock solution is required. For testing with 2 concentrations in singlicate, 850 µl of a 100x stock solution of each concentration is required. For IC50 testing, 1000 µl of  100x stock solution is required. Solid material is also accepted.  

Why profiling with IC50 values?

Kinase panel profiling with IC50 values example with Sorafenib

Example of an FLT3 inhibitor with ³³PanQinase assay: IC50 values of 320 wild type protein kinases are shown with values of lower than 10-4. IC50 value determination for selectivity profiling helps assess differences in the potency of a given compound and allows a precise judgment on the selectivity of kinase inhibitors. 
Only the targets with the lowest IC50 values are named. 

IC50 based profiling is designed to: 

  1. Obtain an accurate guiding parameter for optimization of kinase selectivity during the medicinal chemical optimization phase 
  2. Ensure that observed functional effects of a kinase inhibitor are not due to the inhibition of off-target-kinases and to identify potential adverse side effects of the drug
  3. Provide accurate comparison to a reference compound