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Analysis Biomolecular Interactions of GQDs and Protein by BLI (CAT#: STEM-MB-0121-CJ)

Introduction

Ubiquitin is a small signaling protein with 76 amino acids highly conserved throughout eukaryotes,which is ubiquitously involved in all aspects of cellular activities, such as proteasomal degradation, immune response, and DNA damage repair. It has also been a target protein for studying the interaction involving nanomaterials because of its small size and having one of the lowest molecular weight among the functional proteins. As for graphene quantum dots (GQDs), they have attracted intensive interest due to their extraordinary properties, and great potentials for the development of bio-applications in bioimaging, drug carrier, and so on.




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: Fortebio Bio-Layer Interferometry (BLI)
• Sample Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules
• Optionals: High Resolution Transmission electron Microscope
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