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Analysis of Reactive oxygen species (ROS) by Electron paramagnetic resonance (EPR) spectroscopy (CAT#: STEM-MB-1017-WXH)

Introduction

Reactive oxygen species (ROS) play important roles in cell signalling and homeostasis and are natural by-products of the normal oxygen metabolism. Nevertheless, unfavourable conditions, e.g. environmental stress, can lead to dramatically increased ROS levels that – due to the high reactivity of these molecules – may result in significant damage to the cell. Increased ROS levels are often caused by diseases, for example in inflammatory responses including cardiovascular disease and ischemic injury including stroke and heart attack. Furthermore, ROS are according to the free-radical theory thought to be a major contributor to the organism’s functional decline in the course of aging, rendering their investigation to be of utmost importance.




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.

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