For adapting to competitive environments, organisms, such as viruses and bacteria, often produce toxins and pathogens that target cellular membrane components. Even pharmaceutical products are frequently designed to interact with membrane constituents for better efficacy. The activity of these compounds depends significantly on membrane characteristics, such as membrane composition, surface charge, elasticity, permeability, etc. In turn, exposure to extracytosolic agents may alter membrane assembly, maintenance, and/or function. To acclimate to these induced stresses, cells activate distinct responses commonly controlled by transmembrane signaling, furthering changes in membrane properties. Given how important membrane components are for regulating membrane interactions, it is crucial to study them in isolation from internal cellular processes to understand how external stressors impact specific membrane properties.Model membrane systems, such as solid-supported lipid bilayers (SLBs), are widely used to simulate cellular membranes under controlled conditions. However, most models, utilizing one or more synthetic lipids, cannot capture the intrinsic molecular diversity, limiting their application. To overcome this, the Daniel group has pioneered SLB formation using vesicles extracted from mammalian plasma membranes or outer membranes of Gram-negative bacteria. Membrane vesicles (MVs) provide a realistic model of the native membrane enabling the application of vesicle-derived SLBs to assess membrane biophysics and integrity. Even though considerable efforts have been made in developing SLB platforms showcasing their potential in monitoring subtle changes in membrane properties, their application in the investigation of membrane interactions with molecules of special interest is still lacking. In this dissertation, I have extended the established utility of supported bilayer platforms to understand the impact of specific membrane processes on membrane properties using a combination of surface analytical techniques. I employed liposomes along with MVs isolated from different species to develop SLBs for real-time monitoring of changes in membrane properties influencing and/or arising from membrane interactions. This project provides a means to attain insight into the molecular mechanism of membrane-disrupting agents and membrane responses to such disruptions. My findings connect the existing, simple SLB platforms with complex whole-cell assays for studying membrane interactions with outside interferences and inform the development of novel compounds to modulate these interactions.