Nanotechnological advancements have made a great contribution in developing label-free and highly sensitive biosensors. The detection of ultrasmall adsorbed masses has been enabled by such sensors which transduce molecular interaction into detectable physical quantities. More specifically, microcantilever-based biosensors have caught widespread attention for offering a label-free, highly sensitive and inexpensive platform for biodetection. Although there are a lot of studies investigating microcantilever-based sensors and their biological applications, a comprehensive mathematical modeling and experimental validation of such devices providing a closed form mathematical framework is still lacking. In almost all of the studies, a simple lumped-parameters model has been proposed. However, in order to have a precise biomechanical sensor, a comprehensive model is required being capable of describing all phenomena and dynamics of the biosensor. Therefore, in this study, an extensive distributed-parameters modeling framework is proposed for the piezoelectric microcantilever-based biosensor using different methodologies for the purpose of detecting an ultrasmall adsorbed mass over the microcantilever surface. An optimum modeling methodology is concluded and verified with the experiment. This study includes three main parts. In the first part, the Euler–Bernoulli beam theory is used to model the nonuniform piezoelectric microcantilever. Simulation results are obtained and presented. The same system is then modeled as a nonuniform rectangular plate. The simulation results are presented describing model's capability in the detection of an ultrasmall mass. Finally the last part presents the experimental validation verifying the modeling results. It was shown that plate modeling predicts the real situation with a degree of precision of 99.57% whereas modeling the system as an Euler–Bernoulli beam provides a 94.45% degree of precision. The detection of ultrasmall immobilized enzyme molecules over the cantilever surface was achieved using a self-exciting piezoelectric-based microcantilever in the dynamic mode.