UNIT 8.12 Biosynthetic Labeling of Proteins

Abstract

Biosynthetic labeling techniques are commonly used in the study of biochemical properties, synthesis, processing, intracellular transport, secretion, and degradation of proteins. In this unit, protocols are described for biosynthetically labeling many secreted and membrane proteins. The Basic Protocol describes short-term labeling of cells in suspension (30 min to 3 hr) and an alternate protocol describes a modification of this procedure to be used for adherent cells. Alternate protocols for long-term (3 to 24 hr) and pulse-chase labeling of cells are also presented. Label incorporation can be determined as described in a support protocol. The cells used in these protocols can be either from suspension cultures or from single-cell suspensions of spleen or thymus.