Nonradioactive Analysis Of Biomolecules

Highly sensitive systems which are widely used in molecular biological & biomedical laboratories, such as colorimetric, luminescence, fluorescence measuring using antibody-antigen binding or hybridisation, as well as PCR amplification are described in detail.

New techniques and updated protocols for the detection and analysis of biomolecules - proteins, glycoproteins and nucleic acids. The second edition of this successful laboratory manual describes in detail the highly sensitive systems which are widely used in molecular biological and biomedical laboratories, such as colorimetric, luminescence, fluorescence measuring using antibody/antigen binding or hybridisation as well as PCR amplification. The clearly structured step-by-step protocols with practical hints and a troubleshooting guide are complemented by chapters on the theoretical background and the application of the techniques, enabling scientists to plan, design and conduct the appropriate procedures.

All the bioanalytical labeling and detection techniques in one source! This book gathers together all the important nonradioactive labeling techniques for nucleic acids, proteins, glycoproteins and glycolipids like Digoxigenin:Anti-Dioxigenin (DIG), Biotin, 5-Bromodeoxyuridine (BrdU), Sulfone, Immunogold, Silver Enhancement, and Synthetic Nucleic Acid Probe (SNAP) as well as the standard procedures for optical, chemical, biological, electrochemiluminescent and fluorescent detection. Additionally, applications for the use of non-isotopically labeled biomolecules are described. Specific protocols are given for hybridization analysis such as blot, colony/plaque and in-situ hybridization formats, quantitative formats, and also nonradioactivetechniques for nucleic acid sequencing and amplification. Each chapter contains a short introduction, a detailed description of the method with labprotocols, troubleshooting tips and references.

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

This book combines the experience of 225 experts on 900 pages. Scientists worldwide are currently overwhelmed by the ever-increasing number and diversity of genome projects. This handbook is your guide through the jungle of new methods and techniques available to analyse gene expression - the first to provide such a broad view of the measurement of mRNA and protein expression in vitro, in situ and even in vivo. Despite this broad approach, detail is sufficient for you to grasp the principles behind each method. In each case, the authors weigh up the advantages and disadvantages, paying particular attention to the automated, high-throughput processing demanded by the biotech industry. Completely up to date, the book covers such ground-breaking methods such as DNA microarrays, serial analysis of gene expression, differential display, and identification of open reading frame expressed sequence tags. All the methods and necessary equipment are presented visually in more than 300 mainly colour illustrations to assist their step-by-step reproduction in your laboratory. Each chapter is rounded off with its own set of extensive references that provide access to detailed experimental protocols. In short, the bible of analysing gene expression.

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