Stream Methods Based Characterization of Liquid-Liquid Phase Separation
Biomolecular condensates are organelles formed by liquid-liquid phase separation (LLPS) and play key roles in cellular functions such as signal transduction, transcription, translation and stress response. Phase-separated biomolecular condensates are never in true thermodynamic equilibrium. Therefore, information on LLPS kinetics is needed for a better and more comprehensive understanding of the mechanism of biomolecular condensate formation and its biological functions. However, the early stages of LLPS occur very rapidly, on time scales of 100 ms-10 s. Moreover, the LLPS pathway is very sensitive to experimental conditions and different stimuli (pH jump, urea dilution, salt dilution and protein label cleavage) inducing phase separation can affect the LLPS kinetic trajectory and results.
Customized Services
Our team of experts is very interested in all aspects of LLPS kinetics, including nucleation, growth and transition steps between liquid, gel and aggregates of the state of matter. In the face of the aggregation propensity of phase-separated proteins and the extreme uncertainty of replicating nucleation-dependent kinetic schemes, CD BioSciences offers advanced stream-based methods to help customers study LLPS kinetics of any aggregation-prone protein.
Stopped-Flow Experiment
In the early stages of LLPS, we offer stop-flow experiments to observe such a rapid process. Our cutting-edge stop-flow device allows automated preparation of samples for analysis using relatively small volumes of two, three or four solutions mixed at high speed. It also allows thermal sensing of LLPS and analysis of the thermal regulation of the process.
Capillary Flow Experiment
We offer a detailed, fully automated, high-throughput analysis of LLPS using Capillary Flow Experiment (Capflex). The method allows determination of dilute phase concentration, relative droplet size distribution, kinetics of droplet formation, maturation of concentrated proteins into amyloidogenic fibrils, and binding affinity between LLPS-passed peptides and LLPS-modulated compounds.
Microfluidic Experiment
We offer a more economical microfluidic approach to characterize LLPS, including distinguishing between liquid droplet and solid aggregate formation, studying protein phase transition kinetics, and even determining the rheological properties of condensates. This approach allows manipulation of various parameters of the solution such as protein/RNA concentration, temperature, pH, ionic strength, and solution composition.
Our advanced stream-based methods are widely used to study the complete kinetic trajectory of LLPS under easily controlled near-natural conditions. Depending on the merits of the different methods, we will design customized flow-based solutions for your liquid-liquid phase separation characterization. If you have any special requirements for our services, please feel free to contact us. We are looking forward to working together with your attractive projects.
For research use only, not intended for any clinical use.
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