The ability to analyze the biochemical properties of human cardiac tissue is critical both to an understanding of cardiac pathology and also to the development of novel pharmacotherapies. However current strategies for tissue procurement are not uniform and are potentially biased. In this study we contrasted several commonly used approaches for tissue sampling in order to determine their impact on contractile protein biochemistry. Not surprisingly our results show that different tissue handling strategies have the potential to produce a wide variation in the phosphorylation and proteolysis of selected contractile proteins. However this was not uniform: phosphorylation of troponin I (TnI) and myosin light chain 2 (MLC2) varied significantly depending on approach whereas changes in desmin and myosin binding protein C (MyBP-C) were relatively unaffected. Moreover, some strategies increased whereas others reduced TnI phosphorylation, suggesting a dynamic balance between kinase and phosphatase activities. Overall, procurement strategies that involved maintenance of tissue in cardioplegia solution deviated most dramatically from prompt and rapid tissue immersion in liquid nitrogen.