This is what you expect a clean enzymatic reaction to look like.
Continuous data confirm stable kinetics and reliable rate determination — with 60+ timepoints per replicate instead of one.
The continuous trace exposes experimental noise likely caused by compound insolubility — something a single endpoint could never flag.
Detecting such issues early prevents false conclusions and wasted follow-up.
The curve flattens over time, revealing possible substrate depletion, product inhibition, or slow off-rate behavior.
Early detection of time-dependent inhibition (TDI) provides key clues to mechanism of action (MOA) and compound reversibility.
The lag before steady-state activity hints at conformational effects — for example, the inhibitor preferring an inactive or oligomeric kinase form.
These kinetic patterns are invisible to endpoint assays but critical for mechanistic discovery.
