Here, we present a protocol to isolate the plasma membrane, cytoplasm and mitochondria of U937 cells without the use of high-speed centrifugation. This technique can be used to purify subcellular fractions for subsequent examination of protein localization via immunoblotting.
McCaig, W. D., Deragon, M. A., Haluska Jr,, R. J., Hodges, A. L., Patel, P. S., LaRocca, T. J. Cell Fractionation of U937 Cells in the Absence of High-speed Centrifugation. J. Vis. Exp. (143), e59022, doi:10.3791/59022 (2019).