In experimental animal studies, control sham groups are essential to reduce the influence of the surgical intervention on the analysis. The intraluminal filament procedure is one of the most common models of middle cerebral artery occlusion (MCAO) used in the study of brain ischemia. However, in these studies, the sham group has not usually been included in the experimental design. In this study, we aimed to evaluate the relevance of the sham group by analyzing and comparing the brain protein profile between a sham and an MCAO ischemic group. In the sham group, 98 dysregulated proteins were detected compared to the 171 in the ischemic group. Moreover, a comparative study of both protein profiles showed the existence of a pool of 57 proteins that appeared dysregulated in both sham and ischemic animals. These results indicate that the surgical procedure required for intraluminal occlusion of the MCA induces changes in brain protein expression that are not associated with the ischemic lesion. This study highlights the importance of including control sham groups in the experimental design to guarantee that the therapeutic target under study is not affected by the surgical intervention.