Pulse-chase analysis to measure protein degradation

Abstract

In order to measure protein half-life by pulse-chase analysis, cells are incubated with labeled precursors, [(35)S]-methionine and [(35)S]-cysteine, for a short time (pulse), after which they are washed from the labeled compound and incubated with excess cold precursor (chase). The decay of the amount of protein labeled during the pulse period is measured during the chase with cold precursor.

Keywords: Cellular proteins; Cytoplasmic pools; Derivation of protein half-life; Immunoprecipitation; Methionine pulse-chase; Protein degradation; Pulse-chase analysis.