Over 1000 different lipid species are found in eucaryotic cells with 5 % of our genes solely used for their synthesis [1]. Early discoveries showed that lipids are not only static barriers, used to segregate and compartmentalize the cell but also influence the structure and function of proteins, especially membrane proteins. Lipids function as signaling molecules. To control the lipids' signaling functions, their interaction with effector proteins is regulated via temporal or spatial segregation. Indeed, each membrane leaflet is marked by a distinct lipid composition varying in lipid headgroups and acyl chains [2, 3, 4]. The lipid composition can be influenced by passive effects, like biophysical membrane properties and vesicular transport. In addition, two major protein-mediated lipid distribution systems exist, driven by proteins involved in local lipid synthesis or lipid transport proteins (LTPs) [5,6]. Local lipid synthesis proteins outside the endoplasmic reticulum are often involved in synthesizing low-abundance, transient signaling lipids via lipid precursors. The signaling function of phosphatidylinositol phosphates (PIPs), for example, is tightly controlled by phosphatases and kinases at the plasma membrane [7]. LTPs on the other hand, allow the quick exchange of existing lipid species between membranes or membrane leaflets [5]. LTPs can transport specific lipids with directionality, like flippases and floppases, that use ATP to transport lipids between membrane leaflets against a concentration gradient nurturing the membrane’s asymmetry [8,9]. Others allow the transport of lipids nonspecifically following a concentration gradient, such as scramblases to break membrane asymmetry for signaling purposes [10]. One example is the exposure of phosphatidylserine (PS) to the cell surface by TMEM16F or XKr allowing PS to interact with effector proteins inducing platelet coagulation or apoptosis [11,12]. In most healthy cells, PS levels at the outer leaflet are kept low via the abovementioned flippases such as ATP11A [13]. There is much interest in understanding how lipid dynamics shape protein functions. In this review, we will discuss the available and desirable molecular toolset to study the function of membrane proteins in the context of lipid ligands and the surrounding membrane composition, specifically focusing on ion channels. Ion channels are the second most common drug target and hence our understanding of how lipids impact ion channel function in mammalian cells is imperative.