Tyrosine sulfation is a post-translational modification tied to many biological functions including inflammation, hemostasis, visual functions, and viral infection. In humans, TPSTs are encoded by two genes, TPST1 and TPST2 which catalyzes the transfer of a sulfuryl group from 3’- phosphoadenosine 5’-phosphosulfate. Since the discovery of sulfotyrosine, over 50 proteins containing sites of sulfation have been identified. The role of sulfotyrosine in humans remains a mystery and there is desire to develop workflows to analyze sulfotyrosine containing proteins. Validation methods have not yet been developed for human sulfotyrosine containing proteins identified through bottom-up proteomic analysis. Validation methods will allow for replication of the sulfation modification in vitro and confirm the identified sites are sulfotyrosine. Here we show the methods used to validate a subset of the identified proteins. We proposed two in cellulo methods for validating sulfotyrosine containing proteins: human growth hormone fusion proteins and expressing sulfopeptides as human IgG proteins. Our first method fuses the full-length domain of sulfotyrosine containing proteins to human growth hormone and has the advantage of naturally expressing at high levels in heterologous systems. Our second method fuses tryptic sulfopeptides to the Fc region of human IgG with the advantage of validating sulfotyrosine sites on membrane proteins that would be difficult to purify. Our results demonstrate methods that can be utilized to validate proteins containing the sulfotyrosine site and potentially proteins with other post translational modifications. With the complexity of the human proteome, various methods will be needed to validate the presence of sulfotyrosine in proteins. Our results provide two in cellulo methods. With new validation methods, human sulfotyrosine can continue to be studied and v understood. These methods can be used to replicate the post translational modification in cellulo to allow for subsequent characterization studies of sulfotyrosine containing proteins.