Nonradioactive Labeling And Detection Of Biomolecules

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.

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.

Biological Techniques is a series of volumes aimed at introducing to a wide audience the latest advances in methodology. The pitfalls and problems of new techniques are given due consideration, as are those small but vital details not always explicit in the methods sections of journal papers.In recent years, most biological laboratories have been invaded by computers and a wealth of new DNA technology and this will be reflected in many of the titles appearing in the series.The books will be of value to advanced researchers and graduate students seeking to learn and apply new techniques, and will be useful to teachers of advanced undergraduate courses involving practical or project work.Labelled biomolecules are an essential tool in life science research, and non-radioactive labels are becoming increasingly important due to their convenience of measurement, greater safety and lack of disposal problems compared to radioactive labels.This book provides practical information, background theory and protocols to allow a beginner to label many types of biomolecules, including proteins, peptides, nucleic acids and small molecules.This book is essential for biochemists, molecular biologists and cell biologists wanting to use non-radioactively labelled molecules. Aimed at researchers without specific expertise in chemistry, the book includes: A review of the main signal systems and labels available, indicating their strengths and weaknesses Discussion of the most useful strategies for labelling the various biomolecules 32 protocols covering common labelling needs Descriptions of the factors governing protocol design, enabling protocols to be modified for different applications Sources of information including references, data and suppliers

"Biological Techniques" is a series of volumes aimed at introducing to a wide audience the latest advances in methodology. The pitfalls and problems of new techniques are given due consideration, as are those small but vital details not always explicit in the methods sections of journal papers. In recent years, most biological laboratories have been invaded by computers and a wealth of new DNA technology and this will be reflected in many of the titles appearing in the series. The books will be of value to advanced researchers and graduate students seeking to learn and apply new techniques, and will be useful to teachers of advanced undergraduate courses involving practical or project work. Labelled biomolecules are an essential tool in life science research, and non-radioactive labels are becoming increasingly important due to their convenience of measurement, greater safety and lack of disposal problems compared to radioactive labels. This book provides practical information, background theory and protocols to allow a beginner to label many types of biomolecules, including proteins, peptides, nucleic acids and small molecules.; This book is essential for biochemists, molecular biologists and cell biologists wanting to use non-radioactively labelled molecules. It is aimed at researchers without specific expertise in chemistry, the book includes: a review of the main signal systems and labels available, indicating their strengths and weaknesses. It includes a discussion of the most useful strategies for labelling the various biomolecules. There are 32 protocols covering common labelling needs. There are descriptions of the factors governing protocol design, enabling protocols to be modified for different applications. It contains sources of information including references, data and suppliers.

Protein methods. Biotin labeling methods for proteins and carbohydrates. Direct conjugation of enzymes to proteins. Labeling proteins with fluorochromes. Comparing fluorescent organic dyes for biomolecular labeling. Chemiluminescent detection using alkaline phosphatase-the chemistry. Detection of proteins on filters by enzymatic methods. Enzyme-linked immunosorbent assays. Automation of protein and immunochemistry methods. DNA methods. Biotin labeling methods for DNA. Direct attachment of enzymes to DNA probes. Detection of nucleic acids on filters by enzymatic methods. Polymerase chain reaction and its impact on DNA detection limits. Automated procedures for DNA methods. Tissue and cell methods. Enzyme-based methods in tissue sections. In situ hybridization. Chromosome mapping by fluorescent in situ hybridization. Colloidal gold localization of in situ hybrids. Immunofluorescence methods for microscopic analysis. Flow cytometry methods.

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.