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Analysis Biomolecular Interactions of Oligomeric Variants of GFPp_FGF1 with FGFR1ecd-Fc by BLI (CAT#: STEM-MB-0192-CJ)

Introduction

Fibroblast growth factor receptor 1 (FGFR1) is a receptor tyrosine kinase (RTK) that, together with the extracellular fibroblast growth factors (FGFs), is involved in transmission of signals across the plasma membrane. FGFR1-dependent signaling regulates various biological processes like cell migration, proliferation, and apoptosis. Aberrant activity of this receptor causes many developmental disorders and is detected in numerous cancers. Overexpression of FGFR1 has been observed in various tumor types, like lung, breast, ovarian, prostate, head, and neck cancers. FGFR1 is very efficiently internalized by several endocytic pathways, and thus, its endocytosis can be hijacked for rapid intracellular drug delivery. Importantly, complexes of ligands/targeting molecules with FGFR1 avoid the unfavorable recycling pathway and are predominantly sorted to lysosomes for degradation and cytotoxic drug release. All these features make FGFR1 an attractive molecular target for selective anticancer therapies.




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: FGFR1 (#9740), phospho-FGFR (p-FGFR, #3476), ERK1/2 (#9102), phospho-ERK1/2 (p-ERK1/2, #9101)