•Overview of characterization techniques for molecularly imprinted polymers.•Several techniques are supported by own results.•Conventional techniques are covered such as EM, particle size analyzers, LC-MS/MS.•Advanced characterization techniques include NMR, TGA, DSC.•Specific characterization techniques exist for MIP on surfaces and sensors. Molecularly imprinted polymers (MIP) represent synthetic recognition elements capable of interacting with a target analyte to obtain selective binding and/or detection and/or removal thereof. Various characterization techniques have been explored and evaluated. A first group concerns morphological characterization methods such as electron microscopy (EM), particle size analysis and pore size determination. Secondly, techniques have evolved for analysis of chemical structure characteristics (nuclear magnetic resonance, infrared investigation) and thermal features (thermogravimetric analysis, differential scanning calorimetry). Additionally, selective target binding is mostly studied by liquid chromatography coupled to different detectors. Finally, MIP structures on surfaces such as immobilized particles and films are frequently investigated by EM, atomic force microscopy, static contact angle measurements, X-ray photoelectron spectroscopy, quartz crystal microbalance and surface plasmon resonance. The current review provides an overview of the above mentioned MIP characterization techniques. In most cases, the techniques covered are illustrated with data obtained for MIP against ergot alkaloids.