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Protein-surfactant interactions have been studied due to the numerous applications in different industries including hygienic products, cosmetics, pharmaceuticals, and food products. Single surfactant and mixed surfactant systems with protein are of interest to assess the changes in structure and function of protein. Using several different methods to analyze changes in protein structure, surfactant critical micelle concentration (CMC), and protein-bound surfactant, we can understand the interactions between the surfactant and protein molecules. In this study, two different types of surfactants sodium dodecyl sulfate (SDS) and lauryldimethylamine oxide (LDAO) are analyzed to understand their impact on the protein β-lactoglobulin (βLG). The impact of mixtures of the two surfactants on βLG was also explored. SDS has a significant impact on the structure of βLG at low concentrations. In contrast, higher concentrations of LDAO are required to impact the structure of βLG. The mixtures of the two surfactants with the protein showed minimal changes in surfactant aggregation concentration compared to the mixed surfactant solution with no protein. The mixed surfactant system with protein also required a higher concentration of surfactant to impact the protein structure.