Argonaute 2 (Ago2) is an RNA binding protein that possesses RNase III activity and is a key component of the microRNA pathway in which it cleaves double-stranded RNA. Ago2 is required for essential cellular processes and plays a role in dictating translation of proteins. Commercial antibodies have been utilized for the detection of human and mouse Ago2 using immunohistochemistry (IHC) and western blotting; however, data describing detection in additional mammalian species are currently lacking. Therefore, we evaluated the use of a commercial polyclonal antibody in detecting Ago2 in bovine and porcine tissues. Tissue samples were collected at slaughter and either flash-frozen for western blotting or fixed in 10% neutral buffered formalin and embedded in paraffin for IHC. When bovine and porcine tissue extracts were analyzed by western blotting, anti-Ago2 detected a predominant protein band of ∼108 kDa which closely corresponds to the predicted molecular weight for Ago2; however, multiple bands of lesser intensity and molecular weight were also detected. Using the same antibody, fluorescent IHC on paraffin sections resulted in positive staining for both bovine and porcine tissues including kidney, liver, pancreas, and spleen. Staining for Ago2 was localized in the cytoplasm and appeared to be cell type-specific, with higher signal intensity in endothelial cells, leukocytes, and endocrine cells.