This paper focuses on the influence of thermal history on the rheological properties of unmodified and polymer modified bitumen (PMB), measured at elevated service temperatures, and contributes to the development of test methods for measuring binder properties, which can be used as indicators for asphalt rutting. It was found that the storing and preparation conditions prior to the rheological measurement can have a large influence, especially in the range of long loading times or low frequencies. For elastomer modification, the homogenization and sample pouring temperature and the corresponding change in microstructure, as revealed by fluorescence microscopy, have a large impact on the rheological measurements. For binders with semi-crystalline modifiers, the storage conditions between sample preparation and testing have the largest impact on the rheological behaviour. This can be related to variations in crystallinity, as shown by calorimetry. The main conclusion from this study is that sample preparation and handling is extremely important for the rheological properties of PMBs. Reproducibility can only be achieved when these conditions are controlled more accurately, especially in binder specification tests for rutting susceptibility.