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Kinasera OÜ

Tel. +372 5331 1111
info@kinasera.com
Kinasera live cell assays for monitoring kinase activity in live cells
As ARC-Photo probes detect the active form of protein kinases, the assay is suitable for studying the activation mechanisms of protein kinases inside living cells, e. g., determining the potency of GPCR agonists and antagonist as the activity of PKA is modulated by G-protein (Gs and Gi) coupled receptors or direct activation of adenylate cyclases. Moreover, the assay can be exploited for the determination of cell penetrating properties and cellular activity of PKA inhibitors.

Applications
  • Monitoring protein kinase acitivity in live cells
  • Testing GPCR agonists and antagonists
  • Testing protein kinase inhibitors


  • Kinasera live cell assays are available in two formats:
  • FRET-based assay (Kinasera-FRET live cell assay)

  • For the monitoring of live cells expressing target protein kinases as fusions with fluorescent proteins.
  • Time-resolved fluorescence-based Kinase-Binding assay (Kinasera-TRF live cell assay)

  • For the monitoring of protein kinase activity in native cell lines.


    Kinasera-FRET live cell assay Kinasera-TRF live cell assay
    Assay setup Live cells, 96-well format Live cells, 96-well format
    Assay volume 40 - 100 microL 40 - 100 microL
    Detection principle F�rster's resonance energy transfer (FRET) Time-resolved fluorescence (TRF) (excitation 337 nm, emission in-between 500 and 700 nm)
    Ligand ARC-Fluo probee ARC-Lum probe
    Suitable for monitoring the activation and inhibition of protein kinases in live cells monitoring the activation and inhibition of protein kinases in live cells
    Targets Cell-permeable inhibitors of PKs
    cAMP accumulation and PKA activation
    G-protein coupled receptor agonists and antagonists
    Cell-permeable inhibitors of PKs
    cAMP accumulation and PKA activation
    G-protein coupled receptor agonists and antagonists
    Currently tested for PKA activation (Forskolin) and inhibition (H89) PKA activation (Forskolin, Isoproterenol) and inhibition (H89, Staurosporine)
    Main benefits
  • Applicable for real-time measurements in live cells expressing a PK fused with a fluorescent protein
  • Less affected by low affinity binding protein kinases
  • Applicable for real-time measurements in live cells
  • Applicable in native cells