FLEXYTE® - Protein Kinase Assay Platform

Universal S/T kinase assay

Three generic substrates (KS1, KS2, KS3) enable FLEXYTE® screen assays to be configured for a broad panel of Ser / Thr kinases with performance comparable to radiometric assays.

ATP Km and Substrate Km values for a representative panel of FLEXYTE® kinase assays using the three generic substrates.  Values comparable to those obtained with (γ33P) radiometric assays.

FLT protein kinase assays now developed for: PKBα, PKBβ, PKBγ, p70s6K, SGK1, MSK1, PKA, ROCKII, CHK1, MAPKAPK2, PRAK1 

 

Assay Platform

FLEXYTE® vs Radiometric Assays

Flexlyte vs Radiometric

  • Kinetic parameters determined using FLEXYTE® protein kinase assays are in good agreement to those measured using radiometric methods
  • FLEXYTE® assays gave Z’-factors > 0.9 indicating that they are suitable for HTS applications

Inhibitor Screen

Graph

  • FLEXYTE® protein kinase assay shows excellent comparison with gold-standard radiometric assay and TR-FRET for hit finding (Pearson correlations: 0.98 and 0.93 respectively)

Reliable and Robust

  • Examples of the FLEXYTE® assay using the three generic substrates.  IC50 values were determined for enzyme inhibition by Staurosporine.
  • Values obtained are in excellent agreement to literature values reported using radiometric or TR-FRET methods.

Illustration of Assay Performance

  • Three generic FLEXYTE® peptide substrates enable assays to be configured for >100 Ser/Thr protein kinases
  • FLT assays already developed for: PKBα, PKBβ, PKBγ, p70s6K, SGK1, MSK1, PKA, ROCK-II, CHK1, MAPKAP2, PRAK1