The use of mass spectrometry (MS)-dependent protein research is increasing in the cardiovascular sciences. A major reason for this is the versatility of and ability for MS technologies to accommodate a variety of biological questions such as those pertaining to basic research and clinical applications. In addition, mass spectrometers are becoming easier to operate, and require less expertise to run standard proteomics experiments. Nonetheless, despite the increasing interest in proteomics, many non-expert end users may not be as familiar with the variety of mass spectrometric tools and workflows available to them. We therefore review the major strategies used in unbiased and targeted MS, while providing specific applications in cardiovascular research. Because MS technologies are developing rapidly, it is important to understand the core concepts, strengths and weaknesses. Most importantly, we hope to inspire the further integration of this exciting technology into everyday research in the cardiovascular sciences. (Circ J 2016; 80: 1674-1683).