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Analysis Biomolecular Interactions of Dexlansoprazole with LEDGF/p75 by BLI (CAT#: STEM-MB-0187-CJ)

Introduction

The viral replication of HIV-1 depends on cellular co-factors and cellular pathway. Host cell proteins that particularly assist the viral integration have been determined and termed as integration cofactors (INCFs). The lens epithelium-derived growth factor (LEDGF/p75) is the first identified and the best characterized one to date. The interface of protein-protein interaction between LEDGF/p75 and HIV IN is primarily formed by HIV IN binding domain (LEDGF/p75 IBD) in the C-terminal region of LEDGF/p75 with the HIV IN catalytic core domain (IN CCD). Dexlansoprazole is used to treat certain conditions in which there is too much acid in the stomach. It is used to treat erosive esophagitis or "heartburn" caused by gastroesophageal reflux disease (GERD), a condition where the acid in the stomach washes back up into the esophagus.




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; Virology; 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: Fortebio Bio-Layer Interferometry (BLI)
• Sample Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules
• Optionals: Ni-NTA resin, GST resin