Principles and Reactions of Protein Extraction, Purification, and Characterization

ISBN-13: 9780849320347 / Angielski / Miękka / 2004 / 408 str.

Hafiz Ahmed

Opis

Principles and Reactions of Protein Techniques describes the principles and reactions for most techniques of protein extraction, purification, and characterization. The author focuses on the principles and mechanisms of the experimental procedures and describes the reactions involved. He includes useful advice on which techniques to use and when to apply a particular method. The book discusses the advantages and disadvantages of various protein techniques, enabling students and researchers to master the mechanisms behind the protocols. Seven chapters cover most protein techniques in a consistent format that makes the material easy to retrieve.

Recenzje

"Organized in a logical fashion…the most positive features in each chapter are the occasional placement of actual laboratory procedures (called "working procedures") and discussions on the advantages and disadvantages of each technique/method, often presented in tabular format. This comparison of methods and advice on when certain methods should be used is perhaps the most valuable feature of the book."
- Biochemistry and Molecular Biology Education

Spis treści

EXTRACTION OF PROTEIN
Preparation of Buffers for Protein Extraction
Use of Protease Inhibitors in Protein Extraction
Use of Detergents in Protein Extraction
Chemical Lysis for Protein Extraction
Mechanical Lysis for Protein Extraction
Preparation of Extract from Prokaryotes
Extraction of Recombinant Protein from Bacteria
Preparation of Extracts from Yeast
Preparation of Extracts from Eukaryotes
Preparation of Extracts from Plants
Preparation of Membrane Extracts

ESTIMATION OF PROTEIN
Ultraviolet Absorption Methods
Colorimetric Methods
Fluorescent Methods

ELECTROPHORETIC ANALYSES OF PROTEIN
Driving Force of Electrophoresis
Polyacrylamide Gel Electrophoresis (PAGE)
Isoelectric Focusing (IEF)
Two-Dimensional (2-D) Gel Electrophoresis
Western Blotting
Capillary Electrophoresis

PURIFICATION OF PROTEIN
General Consideration and Purification Strategy
Non-Chromatographic Purification of Proteins
Chromatographic Purification of Proteins

ANTIBODIES: STRUCTURES, INTERACTIONS AND PRODUCTION
Immune System and Antibody Response
Structure of Antibodies
Antigen-Antibody Interactions
Production oof Antibodies
Development of Monoclonal Antibodies
Purification of Antibodies

ANTIBODY LABELING, ANTIBODY DETECTION AND IMMUNOCHEMICAL TECHNIQUES
Methods of Labeling Antibodies
Detection of Antibody Binding
Immunochemical Techniques: a Wide Range of Sensitive Assays for Detecting and Quantitating (Semi) Antigen-Antibody Reactions
Immunoprecipitation
Immunohistochemistry
Flow Cytometry

PURIFICATION OF GLYCOPROTEINS AND ANALYSES OF THEIR OLIGOSACCHARIDES
Diagrams and Stereochemistry of Monosaccharides and Oligosaccharides
Purification of Glycoproteins
Release of Oligosaccharides From Glycoproteins
Preparation of Glycopeptides by Proteolytic Cleavage
Estimation of Carbohydrates
Chromatographic Separation and Detection of Sugars
Electrophoretic Separation
Glycan Detection/Differentiation by Lectins
Analyses of Carbohydrate Linkage

O autorze

Hafiz Ahmed PhD, Hafiz Ahmed