Discovery in Practice · Reference library
Binding assays in microplate assays
Explore binding assays through measurement principles, assay guidance and practical observations. Follow the questions that match your experiment.
5 articles
Ratio measurements & binding
A ratio combines channels that may respond differently to the experiment. Start with those measurements, then examine tracer behavior and fitted binding results.
Your fluorescence polarization result is hiding two measurements
A fluorescence-polarization value is calculated from parallel and perpendicular intensity measurements, and the ratio can hide low photon counts or timing mismatch. The article explains how to preserve both channels, interpret intensity and binding curves, and qualify tracer and reader behavior.
Half the molecules and two thirds of the light
Binding can change tracer brightness and shift an apparent Kd while the curve still fits beautifully. See the equations, controls and fluorescence data to keep.
Binding & proximity assays
Choose a readout that fits the interaction you want to study. Explore tracer design, proximity measurements and the conditions that affect apparent binding.
Your fluorescent label is part of the experiment
Choose a fluorescence polarization tracer by matching lifetime to molecular motion, testing the conjugate and checking controls in the actual assay matrix.
One extra carbon can change the binding result
A one-carbon linker change can alter tracer affinity and assay performance. Learn how to separate binding chemistry from your fluorescence polarization window.
Assay method comparisons
Different methods can report different aspects of the same sample. Compare what each format measures before choosing a readout.
Microplate kinase assays: choose the event you need to measure
ADP production, ATP depletion, substrate phosphorylation and target binding are useful measurements. They are not interchangeable evidence of kinase inhibition.