Abstract
The complexity of the cellular environment is difficult to replicate in vitro. Inert macromolecules such as dextran, Ficoll, and polyethylene glycol (PEG) are common in vitro crowders, but by themselves do not reproduce in-cell biomolecular interactions. Dilute cell lysates or lysis buffer replicate sticking trends observed in cells but do not account for crowding. Recently, we demonstrated that a combination of 150 mg/ml Ficoll and 60% lysis buffer could be used as an in vitro mimic that accounts for both steric crowding and non-steric sticking effects on cytoplasmic protein stability and kinetics.
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