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Analysis of Protein filaments by SDSL EPR (CAT#: STEM-MB-1022-WXH)

Introduction

In numerous neurological diseases like AD, PD, Type 2 diabetes or prion diseases protein misfolding processes lead to protein aggregation and finally to the formation of amyloid- and amyloid-like fibrils that represent the pathological hallmark of these diseases. The exact nature of the actual cytotoxic species is still quite controversial and consequently detailed structural information for the different forms of protein aggregates and insights into the molecular mechanisms of the misfolding processes is of utmost importance for understanding the mechanisms underlying such diseases.
SDSL EPR has turned out to be one of the most powerful techniques for studying such protein aggregates and revealed valuable information especially for several filamentous aggregates being involved in neurological diseases




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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