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Analysis Biomolecular Interactions of Protein with Liposome by BLI (CAT#: STEM-MB-0086-CJ)

Introduction

The interaction of biological relevant molecules and cell membranes plays a key role in drug discovery, drug screening and development for pharmaceutical applications. Artificial membrane systems, such as liposomes, are widely used models to mimic lipid bilayers of biological cell membranes. They are excellent tools to study the biophysical properties and alterations of membranes since lipid composition, lipid ratio and vesicle sizes can be adjusted according to the requirements of the investigation .




Principle

Bio-Layer Interferometry (BLI) is an optical technique for measuring macromolecular interactions by analyzing interference patterns of white light reflected from the surface of a biosensor tip. BLI experiments are used to determine the kinetics and affinity of molecular interactions. In a BLI experiment, one molecule is immobilized to a Dip and Read Biosensor and binding to a second molecule is measured. A change in the number of molecules bound to the end of the biosensor tip causes a shift in the interference pattern that is measured in real-time.

Applications

Immunology/Inflammation; Pharmacology;

Procedure

1. Detect Buffers and prepare samples. BLI experiments are set up with one molecule immobilised on the surface of the biosensor (load sample) and a second molecule in solution (the analytical sample).
2. Fix the load sample on the biocompatible biosensor while the analytical sample is in solution.
3. The biosensor tip is immersed in the solution so that the target molecule begins to bind to the analysis sample.
4. Set up and run the BLI experiment. Molecules bound to or dissociated from the biosensor can generate response curves on the BLI system; unbound molecules, changes in the refractive index of the surrounding medium or changes in flow rate do not affect the interferogram pattern.
5. Collect and analyse data on the BLI's system.

Materials

• Equipment: Gator® Bio-Layer Interferometry (BLI)
• Sample Ty+E217pe: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules
• Optionals: Lipid Vesicles POPC (1-palmitoyl-2-oleoylsn-glycero-3-phosphocholin), DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine), Chloroform, PAA
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