Isolation and functional characterization of the TRAP/mediator complex
S Malik, RG Roeder - Methods in enzymology, 2003 - Elsevier
Publisher Summary This chapter describes the methods developed for isolating functionally
active preparations of the TRAP/Mediator complex and for testing them in cell-free assay
systems. The TRAP/Mediator complex was initially isolated from HeLa cells stably
expressing FLAG-tagged thyroid hormone receptor (TRα). The propagation of this cell-line in
the presence of the cognate ligand (thyroid hormone), but not in its absence, allowed the
isolation (by subsequent affinity chromatography utilizing the FLAG tag) of a multiprotein …
active preparations of the TRAP/Mediator complex and for testing them in cell-free assay
systems. The TRAP/Mediator complex was initially isolated from HeLa cells stably
expressing FLAG-tagged thyroid hormone receptor (TRα). The propagation of this cell-line in
the presence of the cognate ligand (thyroid hormone), but not in its absence, allowed the
isolation (by subsequent affinity chromatography utilizing the FLAG tag) of a multiprotein …