Analysis Biomolecular Interactions of His-Spindlin1 (49–262) and biotin-H3 (1–23) K3me3 by BLI (CAT#: STEM-MB-0225-CJ)

Introduction

Spindlin1 is a methyl-lysine unwinding protein that can have trimethylated lysine in histone H3 through only the second high affinity recognition site 4 (H3K4me3) of the three Tudor-like structural domains. spindlin1 is also an overexpressed gene fragment found in human spindlin1 in ovarian cancer cells as a potential target for drug development. In addition, spindlin1 is highly expressed in various types of malignant tissues, including non-small cell lung cancer, ovarian tumours and some hepatocellular carcinomas. abnormal expression of spindlin1 results in delayed mid-cell cycle and leads to chromosomal instability. Reducing Spindlin1 protein levels strongly impairs the proliferation of liposarcoma cells and increases apoptosis. As the Spindlin1 example shows, methyl-lysine readers have great potential as targets for drug development.

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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: E. coli pellets

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