The human metabolome contains diverse molecules capable of modifying biological macromolecules such as proteins, lipids or nucleic acids without enzymatic assistance, forming non-enzymatic covalent modifications (NECMs). NECMs can cause detrimental effects and are linked to pathologies such as cancer. Among cellular macromolecules, the core histone proteins, which have long half-lives and unstructured nucleophilic tails, are particularly susceptible to NECMs. Recently, we and others have shown that histone glycation adducts caused by the glycolytic byproduct methylglyoxal (MGO) directly affect chromatin structure and function. Moreover, the multifunctional oncoprotein DJ-1 was found to remove early MGO adducts via its deglycase activity. Unfortunately, later-stage histone glycation adducts derived from MGO and other reactive sugars such as ribose, remain difficult to track. Furthermore, uncoupling DJ-1’s deglycase activity from its other functions requires as-yet unavailable chemical tools. Here, I describe the design, synthesis, and evaluation of an alkyne MGO analog (AlkMGO) and a pair of azidoribose (AR) probes to track and enrich glycation modifications, as well as a novel high-throughput-amenable fluorescence-based assay to track DJ-1 activity. Together, these efforts have enabled further study of the physiological effects of glycation and may accelerate the development of small-molecule DJ-1 antagonists.