Bioanalytics: Analytical Methods and Concepts in Biochemistry and Molecular Biology 1st edition by Friedrich Lottspeich, Joachim Engels ISBN 3527694463 97

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ISBN 10:       3527694463
ISBN 13: 978-3527694464
Author:        Friedrich Lottspeich, Joachim W. Engels

 Analytical methods are the essential enabling tools of the modern biosciences. This book presents a comprehensive introduction into these analytical methods, including their physical and chemical backgrounds, as well as a discussion of the strengths and weakness of each method. It covers all major techniques for the determination and experimental analysis of biological macromolecules, including proteins, carbohydrates, lipids and nucleic acids.

The presentation includes frequent cross-references in order to highlight the many connections between different techniques. The book provides a bird’s eye view of the entire subject and enables the reader to select the most appropriate method for any given bioanalytical challenge. This makes the book a handy resource for students and researchers in setting up and evaluating experimental research. The depth of the analysis and the comprehensive nature of the coverage mean that there is also a great deal of new material, even for experienced experimentalists.

The following techniques are covered in detail:

– Purification and determination of proteins
– Measuring enzymatic activity
– Microcalorimetry
– Immunoassays, affinity chromatography and other immunological methods
– Cross-linking, cleavage, and chemical modification of proteins
– Light microscopy, electron microscopy and atomic force microscopy
– Chromatographic and electrophoretic techniques
– Protein sequence and composition analysis
– Mass spectrometry methods
– Measuring protein-protein interactions
– Biosensors
– NMR and EPR of biomolecules
– Electron microscopy and X-ray structure analysis
– Carbohydrate and lipid analysis
– Analysis of posttranslational modifications
– Isolation and determination of nucleic acids
– DNA hybridization techniques
– Polymerase chain reaction techniques
– Protein sequence and composition analysis
– DNA sequence and epigenetic modification analysis
– Analysis of protein-nucleic acid interactions
– Analysis of sequence data
– Proteomics, metabolomics, peptidomics and toponomics
– Chemical biology

Bioanalytics: Analytical Methods and Concepts in Biochemistry and Molecular Biology 1st Table of contents:

Part 1: Protein Analytics

Chapter 1: Protein Purification

• Properties of Proteins: Basic biochemical properties.
• Purification Strategy: Different approaches to isolate proteins.
• Techniques: Methods like homogenization, precipitation, centrifugation, concentration, and detergent removal.
• Sample Preparation for Proteome Analysis: Preprocessing proteins for detailed analysis.

Chapter 2: Protein Determination

• Quantitative Determination: Staining tests and spectroscopic methods for protein quantification.
• Radioactive Labeling: Using radioactive isotopes for labeling peptides and proteins.

Chapter 3: Enzyme Activity Testing

• Catalysts and Enzymes: Study of enzymes as biological catalysts.
• Michaelis-Menten Theory: Concepts like Km and Vmax and their determination.
• Inhibitors: Understanding how inhibitors affect enzyme activity.

Chapter 4: Microcalorimetry

• Techniques like Differential Scanning Calorimetry (DSC), Isothermal Titration Calorimetry (ITC), and Pressure Perturbation Calorimetry (PPC).

Chapter 5: Immunological Techniques

• Antibodies and Antigens: Understanding immune responses and antigen-antibody interactions.
• Antibody Production: Methods for creating antibodies for experimental use.

Chapter 6: Chemical Modification of Proteins and Protein Complexes

• Methods to modify proteins, introduce reporter groups, and crosslink proteins for studying interactions.

Chapter 7: Spectroscopy

• Spectroscopic Techniques: UV/VIS, fluorescence, infrared, Raman spectroscopy, and methods using polarized light.

Chapter 8: Light Microscopy Techniques – Imaging

• Basic Principles: Microscopy evolution, physical principles, and modern applications.
• Fluorescence Microscopy: Special applications and detection methods.

Chapter 9: Cleavage of Proteins

• Proteolytic Enzymes: Breaking down proteins using enzymes.
• Chemical Fragmentation: Approaches like denaturation and alkylation.

Chapter 10: Chromatographic Separation Methods

• Chromatography: Fundamental principles and techniques for separating peptides and proteins, including multidimensional HPLC.

Chapter 11: Electrophoretic Techniques

• Gel Electrophoresis: Theoretical foundations, procedures, and high-resolution two-dimensional electrophoresis.

Chapter 12: Capillary Electrophoresis

• Overview of the basic principles and techniques for electrophoresis using capillary systems.

Chapter 13: Amino Acid Analysis

• Techniques like chromatography and mass spectrometry for analyzing amino acids.

Chapter 14: Protein Sequence Analysis

• Edman Degradation: N-terminal sequence analysis technique.

Chapter 15: Mass Spectrometry

• Ionization and Fragmentation: Understanding mass spectrometry, ion analyzers, and LC-MS for protein and peptide analysis.

Chapter 16: Protein–Protein Interactions

• Techniques like the Two-Hybrid System, co-immunoprecipitation, FRET, and surface plasmon resonance.

Chapter 17: Biosensors

• Applications: From diabetes test strips to biochips, biosensors are used to detect biological substances.

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Part II: 3D Structure Determination

Chapter 18: Magnetic Resonance Spectroscopy of Biomolecules

• NMR and EPR spectroscopy for studying molecular structures.

Chapter 19: Electron Microscopy

• Transmission Electron Microscopy: Techniques for high-resolution biological imaging.

Chapter 20: Atomic Force Microscopy

• AFM: A method to study the surface properties of proteins and biological samples.

Chapter 21: X-Ray Structure Analysis

• Methods like X-Ray Crystallography and SAXS to determine protein structures.

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Part III: Peptides, Carbohydrates, and Lipids

Chapter 22: Analytics of Synthetic Peptides

• Techniques for analyzing synthetic peptides.

Chapter 23: Carbohydrate Analysis

• Methods for analyzing protein glycosylation and the glycome.

Chapter 24: Lipid Analysis

• Techniques for lipid extraction and analysis, including lipidome analysis.

Chapter 25: Analysis of Post-translational Modifications

• Techniques for analyzing phosphorylation and acetylation of proteins.

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Part IV: Nucleic Acid Analytics

Chapter 26: Isolation and Purification of Nucleic Acids

• Methods for purifying DNA and RNA from different sources.

Chapter 27: Analysis of Nucleic Acids

• Restriction Analysis, Electrophoresis, Blotting, and LC-MS of nucleic acids.

Chapter 28: Hybridization and Detection of Nucleic Acids

• Probes and detection systems for hybridization-based techniques.

Chapter 29: Polymerase Chain Reaction (PCR)

• Applications of PCR and other amplification methods.

Chapter 30: DNA Sequencing

• Techniques for Gel-Supported and Next-Generation DNA Sequencing.

Chapter 31: Analysis of Epigenetic Modifications

• Methods for detecting DNA methylation and chromatin modifications.

Chapter 32: Protein–Nucleic Acid Interactions

• Techniques for studying DNA-protein and RNA-protein interactions.

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Part V: Functional and Systems Analytics

Chapter 33: Sequence Data Analysis

• Bioinformatics tools for analyzing sequences and functional predictions.

Chapter 34: Analysis of RNA Transcription

• Methods for studying RNA synthesis and promoter activity.

Chapter 35: Fluorescent In Situ Hybridization

• FISH and CGH applications in molecular cytogenetics.

Chapter 36: Genome Mapping

• Techniques for mapping genetic markers and physical mapping of genomes.

Chapter 37: DNA-Microarray Technology

• Applications in RNA and DNA analysis for functional genomics.

Chapter 38: Oligonucleotides in Cell Biology

• Applications of RNA interference and CRISPR/Cas9 in gene editing.

Chapter 39: Proteome Analysis

• Top-down and Bottom-up proteomics methods for studying the proteome.

Chapter 40: Metabolomics and Peptidomics

• Analyzing metabolites and peptides to understand cellular function.

Chapter 41: Interactomics

• Techniques like protein microarrays to study protein-protein interactions.

Chapter 42: Chemical Biology

• Chemical genetics and approaches to modulate protein function using small molecules.

Chapter 43: Toponome Analysis

• Using antibody-based analysis to study spatial arrangements of proteins and interactions.

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