Amphipol based Membrane Protein Incorporation Service

Although solubilization and stabilization of membrane proteins by detergents is the most general and widely applied approach, there can be risks that use of detergents may cause protein misfolding, loss of stability, and activity. To overcome these problems, a novel class of short amphipathic polymers amphipols (APols) are designed to tightly bind membrane proteins and keep them soluble and stable in more native state. Among various kinds of amphipols, the most commonly used member for membrane protein studies is A8-35, with very high-water solubility (>200 g/L). It is synthesized from low MW polyacrylic acid precursors by forming amide bonds between the carboxylic groups and octylamine or isopropylamine. Although its detergency is too weak to solubilize most membrane proteins, Amphipol has been very successfully used to replace the detergent after the solubilization and stabilize the extracted proteins in their native state in detergent-free solutions.

Amphipols (Creative Biolabs)

Membrane Protein Incorporation in Amphipols

Membrane proteins are usually extracted from membranes using detergents and subsequently transferred to amphipol solutions. When added to the micellar solution at neutral or alkaline pH, amphipol is either mixed with detergents in micelles or binding on the hydrophobic surface of membrane proteins. This way, membrane proteins will be wrapped in small amphipol-detergent complexes. Afterwards, detergent components can be removed by incubation with polystyrene beads.

Membrane Protein Incorporation in Amphipols (Creative Biolabs)

Advantages of Membrane Protein Incorporation in Amphipols

Universal high solubility to keep membrane proteins soluble in aqueous solutions.
Reducing inactivation by the detergent and preserving membrane protein native structure.
Avoiding perturbations of membrane protein's function by detergents. In the vast majority of cases, trapping by Amphipol A8-35 does not affect ligand/substrate binding.
Mild surfactant which provides a favorable environment for proteins to fold or refold from denatured state.
Easily labeled. Differently modified amphipols can be mixed to form multi-functional proteins.

Applications of Membrane Protein Incorporation in Amphipols

Stabilization membrane proteins in native state in detergent-free solutions
Functional studies for ligand/substrate binding
Protein structure with Cryo-EM and NMR
Folding or refolding from denatured state
Immobilization of membrane proteins onto solid supports

Creative Biolabs has been dedicated to providing first-class membrane protein production service using a variety of strategies. Based on our leading-edge platform, we have successfully produced, purified, stabilized, and characterized many challenging membrane protein targets. With the spirit of progress and dedication, we now provide the innovative service of membrane proteins reconstituted into amphipols for various research applications and demands. Please feel free to inquire us to get more details.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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