TL;DR
This chapter presents a detailed protocol for performing in vitro LLPS assays, particularly using polyethylene glycol (PEG) and sodium salt as key modulators of phase behavior.
Biomolecular condensates, formed by proteins, nucleic acids, and other macromolecules through liquid-liquid phase separation, enable molecular enrichment and reaction regulation within membraneless microenvironments. As their physical principles have become better understood, condensates are increasingly being incorpor...
Fluorescent probes based on molecular rotors, based on molecular rotors, with fluorescence lifetime imaging microscopy are combined to quantify changes in protein dynamics during phase separation-associated aggregate formation and provide quantitative, spatially resolved insights into structural transitions.
Liquid-liquid phase separation (LLPS) plays a central role in intracellular compartmentalization and gene regulation. Although optogenetic optoDroplet systems are widely used to study LLPS in living cells, the mechanisms underlying the transition from dynamic liquid-like condensates to operationally defined gel-like as...
ExVivo PhaseScan is established as a versatile tool for dissecting the physicochemical and pathological regulation of condensates, with potential to inform therapeutic strategies for diseases driven by aberrant phase transitions.
Living organisms have evolved multienzyme complexes to achieve efficient and spatially ordered metabolic reactions. Recently, liquid-liquid phase separation (LLPS) has emerged as a fundamental organizational principle underlying these natural networks, providing a versatile platform for constructing artificial multienz...
Liquid-liquid phase separation (LLPS) is a fundamental mechanism that drives biomolecular assembly. The concentration of biomolecules within phase-separated condensates is a critical parameter that reflects their physical state, modulates their biological functions, and provides insights into their formation and aging....
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