Activation of the hemostatic mechanism has been described during thrombolytic therapy. This phenomenon has been detected by new methods of assessing hemostatic system function, based on immunoenzymatic or radioimmunoassays. However, these methods are extremely sensitive and, unless they are performed in expert laboratories, carefully following the recommended procedures, they generate in vitro artifacts. A description of these methods is provided, as well as a critical review of the available studies. The correct use of these methods will provide us with an understanding of the complex response of the hemostatic system to pharmacologic thrombolysis.