Analysis Biomolecular Interactions of Aptamers with Recombinant Human CD25 by BLI (CAT#: STEM-MB-0216-CJ)

Introduction

Multimolecular complexes, including ligand–receptor complexes and receptor multimers, play a central role in mediating a range of processes in cancer biology. They can impact cancer cell growth, differentiation, and survival, as well as the interaction between cancer cells and the microenvironment, which includes antitumor immune responses. A prominent example is the regulation of the immune response by the interaction between IL2 with its alpha receptor subunit, IL2Rα (CD25), which will leads to recruitment of additional receptor subunits (IL2Rβ and γ) to mediate activation signals in lymphocytes. It is important of IL2–CD25 complexes on the immune response in a variety of diseases, including cancer.

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Principle Applications Procedure Materials Notes Related Products

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

Oncology & Cancer; 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: Magnetic cell separation kits, Recombinant human soluble CD25

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