Ser / thr

Ser / thr

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

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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

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  • 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

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  • 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
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