Spectrophotometric kinase activity assays

robust assay format for PKA and PKG kinase assays in kinetic mode

Biaffin provides a comprehensive kinase activity analysis by the use of a spectrophotometric assay. The method couples the phosphorylation of substrates by PKA or PKG and the resulting conversion of ATP to ADP with a reaction catalyzed by pyruvate kinase and lactate dehydrogenase in the presence of phosphoenol pyruvate and NADH which restores the pool of ATP in the assay. For each mole of ADP generated one mole of NADH is oxidized to NAD. Due to to a shift in absorbance from the reduced to the oxidized form of NAD the kinase specific activity is determined from the slope obtained by plotting the initial decrement in absorbance with time. The assay will be performed in 96 well format in kinetic mode.

Our service: 

  • kinase activity studies for cAMP dependent PKA and PKG
  • kinetic characterisation of kinase reaction  
  • determination of enzyme activity, quality control of protein purification processes 
  • inhibitor screening of customer derived compounds
  • IC50 determination at variable ATP concentrations
  • PKA holo enzyme activity studies
  • substrate specificity testing

In addition to services, Biaffin offers products for PKA and PKG research on www.proteinkinase.de and a spectrophotometric PKA kinase activity kit for your in-house projects.

BIAFFIN GmbH & Co KG | Heinrich-Plett-Str. 40 | D-34132 Kassel | Germany
Tel.: +49 (0) 561-804 4661 | Fax: +49 (0) 561-804 4665 | info@biaffin.de

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