We report, for the first time, the use of a 13-amino-acid peptide sequence derived from the calcium-binding site of E-cadherin in the fabrication of an electrochemical peptide-based (E-PB) Pb­(II) sensor. The sensing mechanism is analogous to that of previously developed E-PB sensors. Binding of Pb­(II) rigidifies the surface-immobilized and methylene blue (MB)-modified peptide probe, thereby limiting the accessibility of the tethered MB to the electrode surface. This change in probe flexibility results in a reduction in the MB current that is dependent on the target concentration. The sensor behaves as a “signal-off” sensor in alternating current voltammetry and cyclic voltammetry, but it can behave as a “signal-on” sensor in differential pulse voltammetry when a longer pulse width is employed. It is capable of specific detection of Pb­(II) and is selective enough to be employed in realistically complex samples such as diluted tap water, saliva, and urine samples. The detection is fast; signal saturation can be achieved in <60 s. The sensor can also be fabricated on gold-plated screen-printed carbon electrodes, electrode substrates that are ideal for cost-effective analysis of Pb­(II) in real-world settings.