Microcharacterization of proteins / / R. Kellner, F. Lottspeich, H. E. Meyer

• Record Nr.: UNINA9910830665603321
• Autore: Kellner R.
• Pubbl/distr/stampa: Weinheim, Germany : , : WILEY-VCH, , 1999; ©1999
• ISBN: 1-281-84250-8; 9786611842505; 3-527-61396-X; 3-527-61397-8
• Edizione: [2nd ed.]
• Descrizione fisica: 1 online resource (349 p.)
• Disciplina: 547.75046; 572.65
• Soggetti: Proteins - Analysis - Methodology; Peptides - Analysis - Methodology; Amino acids - Analysis - Methodology
• Lingua di pubblicazione: Inglese
• Formato: Materiale a stampa
• Livello bibliografico: Monografia
• Note generali: Description based upon print version of record.
• Nota di bibliografia: Includes bibliographical references at the end of each chapters and index.
• Nota di contenuto: Microcharacterization of Proteins; Contents; Section I: Overview; I.1 Microcharacterization of Proteins; 1 General Aspects; 2 From a Cell to a Protein Sequence; 3 Genome and Proteome; Section II: Microseparation Techniques and Sample Preparation; II.1 Microseparation Techniques I: High Resolution Gel-Electrophoretic Techniques : Qualitative, Quantitative and Micropreparative Applications; 1 Introduction; 2 Theory; 3 Media and Equipment; 3.1 Media; 3.2 Equipment; 4 Gel Electrophoretic Methods; 4.1 Disc Electrophoresis; 4.2 Gradient Gel Electrophoresis; 4.3 Additives in Electrophoresis; 4.4 SDS Electrophoresis4.5 Isoelectric Focusing; 4.6 Two-Dimensional Electrophoresis (2 -DE); 5 References; II.2 Microseparation Techniques II: Gel Electrophoresis for Sample Preparation in Protein Chemistry; 1 Introduction; 2 Denaturing Techniques; 2.1 Commonly Used SDS-Polyacrylamide Gel Electrophoresis Techniques for Protein Separation; 2.2 Blue-SDS-PAGE for Recovery of Membrane Proteins from Gels; 2.3 ; Electroelution of Proteins after Blue-SDS-PAGE; 2.4 Electroblotting of Blue and Colorless SDS Gels; 2.5 Isoelectric Focusing in the Presence of Urea; 3 Native Techniques; 3.1 Colorless-Native-PAGE3.2 Blue-Native-PAGE; 3.3 Native Isoelectric Focusing; 4 References; II.3 Microseparation Techniques III: Electroblotting; 1 Introduction; 2 Electroblotting; 2.1 Polyacrylamide Gel Electrophoresis; 2.2 Blot Systems; 2.2.1 Tank Blotting; 2.2.2 Semidiy Blotting; 2.3 Blotting Parameters; 2.3.1 The Blotting Process; 2.3.2 Transfer Buffers; 2.3.3 Addition of SDS; 2.3.4 Addition of Methanol; 2.3.5 Influence of Protein Concentration; 3 Blotting Membranes; 4 References; II.4 Microseparation Techniques IV: Analysis of Peptides and Proteins by Capillary Electrophoresis; 1 Introduction2 Theory; 2.1 Capillary Isotachophoresis; 2.2 Capillary Zone Electrophoresis; 2.3 Electroosmotic Flow; 3 Instrumentation; 3.1 Injection; 3.2 Detection; 4 Applications; 4.1 Peptide Separations; 4.2 Protein Separations; 5 References; II.5 Microseparation Techniques V: High Performance Liquid Chromatography; 1 Introduction; 2 Principle of HPLC; 3 Getting Started; 3.1 Solvents; 3.2 Pump; 3.3 Gradient; 3.4 Pre-Column Split; 3.5 Sample Preparation; 3.6 Injector; 3.7 Tubings; 3.8 In-Line Filter, Guard Column; 3.9 Column; 3.10 Detection; 3.11 Fractionation; 4 Applications; 5 References; II.6 Sample Preparation I: Removal of Salts and Detergents1 Introduction; 2 Removal of Salts or Polar Components; 2.1 Protein-Binding Membranes; 2.2 High Performance Liquid Chromatography; 2.3 Desalting on Microcolumns; 3 Removal of Detergents and Apolar Contaminants; 3.1 Detergents; 3.2 Protein-binding Membranes; 3.3 Precipitation; 3.4 SDS-PAGE; 3.5 Concentrating Gels; 4 "Golden Rules" for Protein and Peptide Handling; 5 References; II.7 Sample Preparation II: Chemical and Enzymatic Fragmentation of Proteins; 1 Strategy; 2 Denaturation, Reduction and Alkylation; 3 Enzymatic Fragmentation; 3.1 Enzymes
• Sommario/riassunto: Proteomics - the analysis of the whole set of proteins and their functions in a cell - is based on the revolutionary developments which have been achieved in protein analysis during the last years. The number of finished genome projects is growing and in parallel there is a dramatically increasing need to identify the products of revealed genes. Acting on a micro level modern protein chemistry increases our understanding of biological events by elucidating the relevant structure-function relationships. The second edition of the successful title Microcharacterization of Proteins present