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Right here, we provide a formal framework and a web-based device, the SensorOverlord, that predicts the input range of two-state ratiometric biosensors because of the experimental error in calculating their signal. We demonstrate the energy of this tool by predicting the product range of values that can be assessed accurately by biosensors that detect pH, NAD+, NADH, NADPH, histidine, and glutathione redox potential. The SensorOverlord enables users to compare the predicted reliability of