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Analysis Biomolecular Interactions of Cell-free Expressed Lectins with Carbohydrate Ligands by BLI (CAT#: STEM-MB-0094-CJ)

Introduction

Lectins, or hemagglutinins, are an “anti-nutrient” that have received much attention due to popular media and fad diet books citing lectins as a major cause for obesity, chronic inflammation, and autoimmune diseases. Lectins are defined as proteins that bind to carbohydrates. The same features that lectins use to defend plants in nature may cause problems during human digestion. They resist being broken down in the gut and are stable in acidic environments, features that protect lectin-containing plants in nature.




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

synthetic glycobiology

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 Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules