We set out to first establish and validate a novel rapid method of enriching adult Müller cells, which yields sufficient cell numbers for proteomic profiling while keeping cells viable and morphologically, as well as physiologically fully intact. From these preparations we collected a comprehensive data set of the proteome of native murine Müller cells. Since many Müller cell functions depend on cell-cell interactions occurring at the plasma membrane level, we additionally included a subcellular fractionation approach that enabled to discriminate proteins primarily residing in the cytosol or the nucleus from those allocated in the plasma membrane and thus generate a first time Müller cell surfaceome study. We consider these data a first important step to better understanding the complex crosstalk between various retinal cell types and Müller cells as key players in health and ultimately also under disease conditions.