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Analysis of Human islet amyloid polypeptide (hIAAP) by SDSL EPR (CAT#: STEM-MB-1025-WXH)

Introduction

Human Islet Amyloid Polypeptide (hIAPP, amylin) misfolding plays an important role in the pathogenesis of Type 2 diabetes mellitus. The 37-residue peptide is synthesized in pancreatic islet β-cells and cosecreted with insulin. It is the primary amyloidogenic agent in this abundant disease and hIAPP fibrils are found in more than 90% of Type 2 diabetes patients.




Principle

Electron Paramagnetic Resonance (EPR), also called Electron Spin Resonance (ESR), is a branch of magnetic resonance spectroscopy which utilizes microwave radiation to probe species with unpaired electrons, such as radicals, radical cations, and triplets in the presence of an externally applied static magnetic field.
EPR spectroscopy is particularly suitable for the investigation of (bio)chemical systems with strongly localized spin density and their interaction with the environment. For these systems EPR provides information on the structure and dynamics and is widely used in chemistry, physics and biology.
Site-directed spin labeling (SDSL) is a technique for investigating the structure and local dynamics of proteins using electron spin resonance. The theory of SDSL is based on the specific reaction of spin labels with amino acids. A spin label's built-in protein structure can be detected by EPR spectroscopy. SDSL is also a useful tool in examinations of the protein folding process.

Applications

• Study dynamic organisation of lipids in biological membranes, lipid-protein interactions and temperature of transition of gel to liquid crystalline phases.
• Determine oxygen levels in tissues and blood.
• Injection of spin-labeled molecules allows for electron resonance imaging of living organisms.
• EPR can be used to measure microviscosity and micropolarity within drug delivery systems as well as the characterization of colloidal drug carriers.
• The study of radiation-induced free radicals in biological substances (for cancer research).
• Investigation on the antioxidant properties of medicine

Procedure

1. Sample Preparation
2. Electron paramagnetic resonance (EPR) spectroscopy testing
3. Data analysis

Materials

• EPR Spectrometer
• Spectrophotometer
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