Determination of Concentration and Partition Coefficients of Components in Biomolecular Condensates

In general, the interactions between biomolecules and the activity of subsequent biological pathways are closely related to the concentration of their components. Biomolecular condensates enhance biomolecular interactions through liquid-liquid phase separation (LLPS). Therefore, the concentration of internal components is an important material property of biomolecular condensates. LLPS occurs at a certain concentration, and a dynamic equilibrium between the dilute and dense phases is reached, which is called the saturation concentration (C_sat). The ratio of the concentration of the components in the dense phase (C_den) to the concentration of the dilute phase (C_den/C_sat) is made the partition coefficient (K). In a simple binary phase separation system under a specific set of conditions, based on the underlying physics of LLPS, both C_sat and K are fixed.

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Fluctuations in protein and RNA concentrations can cause negative effects, such as biological noise, on a range of cellular processes. However, the formation of biomolecular condensates and the ensuing inherent concentration effects can buffer the noise caused by fluctuations in biomolecular concentrations. Our experts are developing detailed methodological protocols to determine the concentration and partition coefficients of proteins and RNA, which are important material properties of biomolecular condensates. CD BioSciences offers customized determination services of concentration and partition coefficient of components in biomolecular condensates to help you easily obtain the concentration of dense and dilute phases.

We are proud to offer fluorescence correlation spectroscopy (FCS) to determine the concentration and diffusivity of components in biomolecular condensates. This technique is based on fluctuations in fluorescence intensity due to Brownian diffusion of fluorescent molecules and/or physicochemical reactions. Our FCS technique offers the following advantages:

✓Can analyze volumes as small as 10^-15 L, helping you to calculate the hydrodynamic properties and behavior of individual molecules.

✓Performed on a fluorescence correlation microscope, correlation curves can be generated over time from the recorded data, helping you to calculate the concentration, diffusivity and hydrodynamic radius of the biomolecule under study.

✓The excitation zone can be set inside or outside the studied bioconcentrate, estimating the material properties of the dense or dilute phase, respectively.

✓The formation of nanoscale condensates can be captured to obtain the critical concentration for nanoscale phase separation.

✓The size and growth rate of the biocondensate, as well as the composition and binding affinity of the internal biomolecules, can be determined.

✓ It can also be easily performed on living cells.

In addition, we offer experimental measurements (by shake flask, HPLC, etc.) or calculations based on various methods (fragment-based, atomic-based, etc.) to directly estimate partition coefficients.

Our services are widely used to determine the concentration and partition coefficients of components in biomolecular condensates, which can influence the interaction between molecules and the activity of their biological pathways. 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.