Analysis Kinetics of IL-8 Protein by KinExA (CAT#: STEM-MB-0263-CJ)

Introduction

Glioblastomas (GBMs) are the most lethal malignancy in the human tumor. A significant portion of the vascular endothelium has a neoplastic origin, and the glioblastomas stem-like cells (GSCs) have the potential of transdifferentiating to endothelial like cells. This transdifferentiation process involves a series of cytokines, including vascular endothelial growth factor (VEGF), transforming growth factor beta, and interleukin-8（IL-8). IL-8 is a chemokine with a defining CXC motif secreted by many kinds of cells. In human gliomas, IL-8 is expressed and secreted at high levels both in vitro and in vivo. The level of IL-8 is associated with the histological grade of glioma, and the most malignant form of glioblastoma shows the highest expression in necrotic cells, hypoxia conditions may stimulate IL-8 expression. And in a glioma niche, endothelial cells and TDEC can be stimulated by IL-8 in hypoxia condition.

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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

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