PhosphoSens CAMKK1 Protein Kinase Assays, Substrates & Recombinant Enzymes

PhosphoSens peptide substrates are compatible with assay buffers supplemented with defined calcium and calmodulin concentrations, enabling real-time monitoring of CAMKK1 activity under physiologically relevant cofactor conditions. Continuous progress curves make it straightforward to confirm that kinase activation is saturated and that reactions are in a linear regime before compound IC50 measurements are collected. This eliminates the ambiguity inherent in single-endpoint assays where incomplete cofactor activation can produce artifactual results.

Yes. Because PhosphoSens generates continuous fluorescence progress curves, time-dependent changes in enzyme velocity are directly observable, allowing accurate determination of kinact and KI for covalent or slowly binding inhibitors. This is a significant advantage over ADP-Glo or HTRF endpoint assays, which collapse the entire reaction into a single data point and cannot distinguish rapid reversible inhibition from slow covalent engagement. Real-time kinetics are especially valuable as covalent CAMKK1 inhibitor programs emerge.

PhosphoSens assays can be run at physiological ATP concentrations (1-2 mM), which is critical for accurately ranking inhibitor potency in a cellular context. Many competing assay formats use low ATP (10-100 u00b5M) to maximize signal, which artificially inflates the apparent potency of ATP-competitive inhibitors. Using near-physiological ATP with PhosphoSens yields IC50 values that better predict compound efficacy in cells and reduces costly late-stage attrition in CAMKK1 drug discovery campaigns.