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METHODS IN ENZYMOLOGY
Methods in Enzymology
Section 1: Hydrogen Peroxide Detection Methods
Chapter One. Photooxidation of Amplex Red to Resorufin: Implications of Exposing the Amplex Red Assay to Light
2 Amplex Red Assay and Its Possible Artifacts due to Light Exposure
3 Experimental Considerations
Chapter Two. Boronate-Based Fluorescent Probes: Imaging Hydrogen Peroxide in Living Systems
2 Fluorescent Boronate-Based Hydrogen Peroxide Probes
3 Examples of Probes and Their Usage
4 Imaging Endogenous H2O2 Using Fluorescent Probes
Chapter Three. Visualization of Intracellular Hydrogen Peroxide with HyPer, a Genetically Encoded Fluorescent Probe
2 Experimental Design
Chapter Four. In Vivo Imaging of H2O2 Production in Drosophila
4 Image and Data Analysis
5 Data Interpretation
Chapter Five. Single Fluorescent Probe Distinguishes Hydrogen Peroxide and Nitric Oxide in Cell Imaging
2 A Guide to Single Fluorescent Probe Distinguishes H2O2 and NO
3 Fluorescence Imaging of Endogenously Produced H2O2 and NO in RAW 264.7 Macrophages Cells
Chapter Six. Electrochemical Biosensors for On-Chip Detection of Oxidative Stress from Cells
2 Experimental Components and Procedures
3 Methods of Calibrating Enzyme Sensors
4 Monitoring ROS Production from Cells Using Electrochemistry
Chapter Seven. Electrochemical Detection of H2O2 Formation in Isolated Mitochondria
2 The Electrochemical Chamber
3 The Platinized Carbon Fiber Microelectrode
4 Data Analysis for H2O2 Amperometric Measurements
5 H2O2 and O2 Calibration Tests
6 Preparation of Isolated Mitochondria
7 Measure of H2O2 from Mitochondrial Suspension
Chapter Eight. Detection of H2O2 by Fluorescence Correlation Spectroscopy
2 Experimental Methods
3 Comparison of Sensitivity Between the FCS-Based Method and an Amplex Red-Based Method
Chapter Nine. Real-Time Monitoring of Reactive Oxygen and Nitrogen Species in a Multiwell Plate Using the Diagnostic Marker Products of Specific Probes
2 Experimental Methods
3 Results and Discussion
Section 2: Temporal and Spatial H2O2 Distribution
Chapter Ten. H2O2 Delivery to Cells: Steady-State Versus Bolus Addition
2 Experimental Components and Considerations
3 Pilot Experiments
4 Steady-State Experimental Implementation
Chapter Eleven. Imaging H2O2 Microdomains in Receptor Tyrosine Kinases Signaling
2 Experimental Design
6 Anticipated Results
Section 3: Cellular Sources of H2O2
Chapter Twelve. The Determination and Analysis of Site-Specific Rates of Mitochondrial Reactive Oxygen Species Production
2 H2O2 Production Measurements in Isolated Mitochondria
3 Maximum Production Rates from Specific Sites: The Nature and Capacity of the Machinery
4 Concluding Remarks
Chapter Thirteen. A Microfluidic Systems Biology Approach for Live Single-Cell Mitochondrial ROS Imaging
2 Microfluidic Platform
3 Microscope System and Image Analysis
4 Imaging Mitochondrial Superoxide Production
5 Imaging Mitochondrial Hydrogen Peroxide Production
7 Potential Pitfalls
Chapter Fourteen. Detection of Oxidative Damage in Response to Protein Misfolding in the Endoplasmic Reticulum
2 UPR-Induced Oxidative Damage
This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This is the first of three volumes on hydrogen peroxide and cell signaling, and includes chapters on such topics as photooxidation of amplex red to resorufin, boronate-based fluorescent probes, and visualization of intracellular hydrogen peroxide with HyPer.
- Continues the legacy of this premier serial with quality chapters authored by leaders in the field
- Covers hydrogen peroxide and cell signaling
- Contains chapters on such topics as photooxidation of amplex red to resorufin, boronate-based fluorescent probes, and visualization of intracellular hydrogen peroxide with HyPer
Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists.
- No. of pages:
- © Academic Press 2013
- 19th June 2013
- Academic Press
- Hardcover ISBN:
- eBook ISBN:
Lester Packer received a PhD in Microbiology and Biochemistry in 1956 from Yale University. In 1961, he joined the University of California at Berkeley serving as Professor of Cell and Molecular Biology until 2000, and then was appointed Adjunct Professor, Pharmacology and Pharmaceutical Sciences, School of Pharmacy at the University of Southern California.
Dr Packer received numerous distinctions including three honorary doctoral degrees, several distinguished Professor appointments. He was awarded Chevalier de l’Ordre National du Merite (Knight of the French National Order of Merit) and later promoted to the rank of Officier. He served as President of the Society for Free Radical Research International (SFRRI), founder and Honorary President of the Oxygen Club of California.
He has edited numerous books and published research; some of the most cited articles have become classics in the field of free radical biology:
Dr Packer is a member of many professional societies and editorial boards. His research elucidated - the Antioxidant Network concept. Exogenous lipoic acid was discovered to be one of the most potent natural antioxidants and placed as the ultimate reductant or in the pecking order of the “Antioxidant Network” regenerating vitamins C and E and stimulating glutathione synthesis, thereby improving the overall cellular antioxidant defense. The Antioxidant Network is a concept addressing the cell’s redox status. He established a world-wide network of research programs by supporting and co-organizing conferences on free radical research and redox biology in Asia, Europe, and America.
Department of Molecular Pharmacology and Toxicology, School of Pharmaceutical Sciences, University of Southern California, USA
ENRIQUE CADENAS, MD, PhD, received his PhD in biochemistry from the University of Buenos Aires, School of Medicine. He is professor of pharmacology and pharmaceutical sciences at the University of Southern California School of Pharmacy and of biochemistry and molecular biology at the University of Southern California Keck School of Medicine, and doctor honoris causa (medicine) at the University of Linköping, Sweden. Cadenas was president of the Society for Free Radical Research International (SFRRI) and is fellow of the Society for Free Radical Biology & Medicine. He served the scientific community by participating on NIH study sections (2002-2006; chair 2006-2008). His research interests include energy and redox metabolism in brain aging and the coordinated inflammatory-metabolic responses in brain and neurodegenerative diseases.
Pharmacology & Pharmaceutical Sciences, School of Pharmacy, University of Southern California, USA
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