- Protocol for Designing Small-Molecule-Regulated Destabilizing Domains for In Vitro Use.
Protocol for Designing Small-Molecule-Regulated Destabilizing Domains for In Vitro Use.
STAR protocols (2020-10-01)
Prerana Ramadurgum, John D Hulleman
PMID32995752
ABSTRACT
The use of destabilizing domains (DDs) to conditionally control the abundance of a protein of interest (POI) through a small-molecule stabilizer has gained increasing traction both in vitro and in vivo. Yet there are specific considerations for the development and accurate control of user-defined POIs via DDs, as well as the identification of novel (and potentially synergistic) small-molecule stabilizers. Here, we describe a platform for achieving these goals. For complete details on the use and execution of this protocol, please refer to Ramadurgum et al. (2020).
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Ponceau S solution, BioReagent, suitable (for use in cellulose acetate electrophoresis), 0.1 % (w/v) in 5% acetic acid
Sigma-Aldrich
Hanks′ Balanced Salt solution, Modified, with sodium bicarbonate, without phenol red, calcium chloride and magnesium sulfate, liquid, sterile-filtered, suitable for cell culture