S'identifier

Albany College of Pharmacy and Health Sciences

2 ARTICLES PUBLISHED IN JoVE

Biochemistry

Cell Fractionation of U937 Cells in the Absence of High-speed Centrifugation

William D. McCaig¹, Matthew A. Deragon¹, Robert J. Haluska Jr,¹, Alexa L. Hodges¹, Payal S. Patel¹, Timothy J. LaRocca¹

¹Department of Basic and Clinical Sciences, Albany College of Pharmacy and Health Sciences

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.

Biochemistry

Cell Fractionation of U937 Cells by Isopycnic Density Gradient Purification

William D. McCaig¹, Matthew A. Deragon¹, Phillip V. Truong¹, Angeleigh R. Knapp¹, Timothy J. LaRocca¹

¹Department of Basic and Clinical Sciences, Albany College of Pharmacy and Health Sciences

This fractionation protocol will allow researchers to isolate cytoplasmic, nuclear, mitochondrial, and membrane proteins from mammalian cells. The latter two subcellular fractions are further purified via isopycnic density gradient.