Background of Solid Supported Membranes
Solid Support Membrane (SSM) is a convenient biofilm model system where solutions can be rapidly exchanged at the surface. SSM is built on a planar gold electrode and consists of a mixed streptavidin-phospholipid bilayer. Lipoprotein bodies, membrane vesicles or membrane fragments containing the transporter protein of interest are adsorbed onto the SSM surface to form a capacitively coupled membrane system. The SSM behaves like a capacitor, with the solution in a fluid chamber and the gold electrode as the capacitor plate. A rapid jump in substrate concentration activates the protein. Charge migration is then measured by capacitive coupling through the support membrane. SSM-based electrophysiological measurements have been widely used for functional characterization of ion pumps and transporters as a promising platform technology for drug screening and development.
Fig. 1. Adsorption geometry of proteoliposomes on solid supported membranes. (Bazzone A, et al., 2013)
Solid Supported Membranesy-based Lysosomal Channel Research Services
Scientists have tested many different electrogenic proteins (ion pumps, transporter proteins and channels) using SSM electrophysiology. Here, CD BioSciences focuses on applying solid support membrane technology to intracellular lysosomal transporter proteins to overcome the inaccessibility of lysosomal endosomal membranes. Our customized measurement service for lysosomal channels on solid support membranes has been widely used for functional characterization of the following lysosomal ion pumps and transporters.
- Lysosomal CLC-7 Ion Channels
- Lysosomal V-ATPase
Our biologists obtain membrane fractions from natural tissues or lysosomal intracellular membranes rich in plasma membrane or lysosomes in transfected cells by sugar gradient isolation to further measure proteins expressed in intracellular lysosomal ion channels in their natural environment. We develop customized procedural procedures for SSM-based electrophysiological experiments for the measurement of lysosomal channels.
(1) Setting up SSM-based electrophysiology equipment.
(2) Preparing the SSM-based electrophysiology experiments, including the lipid solution used to form the SSM, chlorination of the reference electrode, preparation of the polyacrylamide gel bridge and measurement solution.
(3) Measuring the SSM Electrical Parameters
(4) Examing the solution exchange artifacts.
(5) Adding protein samples.
(6) Concentration jump experiment with signal recording.
Why Choose Us
- No complex set-up procedures are required.
- Particularly useful in situations where conventional electrophysiology (i.e., membrane clamp and voltage clamp methods) cannot be applied.
- Provides a unique opportunity to perform electrophysiological studies with high temporal resolution using arbitrary substrates.
- Native environment.
- Automated.
- Suitable for screening.
CD BioSciences can meet any reasonable needs of our clients, taking time and budget into consideration for you. Our aim is to be customer-centric and to provide the highest quality services to customers. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.
Reference
- Bazzone A, et al. (2013) Introduction to solid supported membrane based electrophysiology. J Vis Exp. (75):e50230.