CHX is mostly used only in in vitro experiments, and is not suitable for human use as a therapeutic compound. The best-known use of CHX is as an experimental tool in molecular biology to determine the half-life
Thus, the CHX chase assay is widely recognized and used to observe intracellular protein degradation and to determine the half-life of a given protein in eukaryotes. Here, we present a complete experimental procedure of the CHX chase assay. Graphical overview...
the level of remaining Drp1 at different time points was quantified as the percentage of initial Drp1 level (0 hour of CHX treatment). C and D, the half-life of Drp1 in the presence of GFP-Parkin or GFP alone was analyzed by [35S] methionine pulse-chase assay, and the protein levels ...
S]methioninepulse-chaseassayfor thehalf-lifeofDrp1.A,293TcellsweretransfectedwithGFP-ParkinandGFP-Parkin(ΔR2) for24hours.CellswerethentreatedwithCHX(100μg/ml)fortheindicatedhours,and Westernblottingwasperformed.B,thelevelofremainingDrp1atdifferenttimepointswas ...
Cycloheximide (CHX) chase assay and [35S] methionine pulse-chase assay for the half-life of Drp1. A, 293T cells were transfected with GFP-Parkin and GFP-Parkin (ΔR2) for 24 hours. Cells were then treated with CHX (100 μg/ml) for the indicated hours, and Western blotting was ...