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Analysis Affinity of IGF-IR to IGF-I and IGF-II by KinExA (CAT#: STEM-MB-0034-CJ)

Introduction

Insulin-like growth factor-I receptor (IGF-IR) plays an important role in cell growth and differentiation. Overexpression of IGF-IR leads to abnormal cell proliferation and malignant transformation. IGF-IR deficient cells are resistant to carcinogenic transformation. In addition to its role in cell proliferation, IGF-IR also plays an important role in tumor cell survival. Overexpressed IGF-IR protects cells from apoptosis induced by serum withdrawal, tumor necrosis factor, etoposide, or interleukin-3 withdrawal in vitro. Elevated plasma levels of IGF-I are associated with prostate, breast, lung, and colon cancer risk. IGF-II is often overexpressed in colorectal cancer, neuroblastoma, liver, adrenal cortex, and Wilms tumors. IGF-IR is a signaling receptor for IGF-I and IGF-II ligands and is also overexpressed in some human cancers. The expression of IGF-IR on many tumor cell types suggests that this anti-IGF-IR monoclonal antibody has the potential to treat many different types of cancer.




Principle

KinExA is a two-stage analysis system. In the first stage, a number of solutions are prepared, where one partner remains constant (constant binding partner, or CBP) and the other (titrant) is variable, usually serially diluted. As the titrant is added, the free CBP decreases and is analysed by a sophisticated and precise microfluorescence measurement device. The signal generated can be mathematically related to the affinity (KD) of the two molecules for each other, as well as the kinetic parameters of binding (kon) and dissociation (koff).

Applications

Oncology & Cancer; Immunology/Inflammation, Toxicology; Pharmacology

Procedure

1. preparation of the functionalized beads which will capture the analyte for measuremen.
2. preparation of a series of solutions consisting of a constant initial concentration of one component of the binary reaction and serial dilutions of the other reactant. The component that is kept constant is the constant binding partner (CBP) , and is the one which will be analyzed.
3. each reaction mixture is sampled and the fluorescence of free CBP bound to the capture beads is obtained for subsequent numerical analysis.

Materials

• Sample Type: Antibodies, Cells, Small Molecules, Proteins, DNA, Lipids, Serum, & More
• Equipment: Kinetic Exclusion Assay (KinExA)
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