Microcharacterization of proteins [[electronic resource] /] / R. Kellner, F. Lottspeich, H.E. Meyer

• Record Nr.: UNISA996197616503316
• Autore: Kellner R (Roland)
• Pubbl/distr/stampa: Weinheim, : VCH, c1994
• ISBN: 1-281-75875-2; 9786611758752; 3-527-61571-7; 3-527-61570-9
• Descrizione fisica: 1 online resource (288 p.)
• Altri autori (Persone): LottspeichF <1947-> (Friedrich); MeyerH. E (Helmut E.)
• Disciplina: 547.75046; 572/.65
• Soggetti: Proteins; Proteins - Analysis
• Lingua di pubblicazione: Inglese
• Formato: Materiale a stampa
• Livello bibliografico: Monografia
• Note generali: Presented at the first meeting "Mikromethoden in der Proteinchemie" held June 1994 at the Max Planck Institute for Biochemistry.
• Nota di bibliografia: Includes bibliographical references.
• Nota di contenuto: Microcharacterization of Proteins; Contents; Section I: Overview; 1.1 Microcharacterization of Proteins; 1 General Aspects; 2 From a Cell to a Protein Sequence; 3 FutureTrends; Section II: Sample Preparation; II.1 Chemical and Enzymatic Fragmentation of Proteins; 1 Strategy; 2 Denaturation, Reduction and Alkylation; 3 Enzymatic Fragmentation; 3.1 Enzymes; 3.2 Practical Considerations; 4 Chemical Fragmentation; 4.1 Cyanogen Bromide Cleavage; 4.2 Partial Acid Hydrolysis; 4.3 Hydroxylamine Cleavage of Asn-Gly Bonds; 4.4 Cleavage at Tryptophan; 4.5 Cleavage at Cysteine; 5 References; II.2 Microseparation Techniques I: High Performance Liquid Chromatography1 Introduction; 2 Getting Started; 2.1 Solvents; 2.2 Pump; 2.3 Pre-Column Split; 2.4 Sample Preparation; 2.5 Injector; 2.6 Tubings; 2.7 In-Line Filter, Guard Column; 2.8 Analytical Column; 2.9 Elution; 2.10 Detection; 2.11 Fractionation; 3 Applications; 4 References; II.3 Microseparation Techniques II: Analysis of Peptides and Proteins by Capillary Electrophoresis; 1 Introduction; 2 Theory; 2.1 ; Capillary Isotachophoresis; 2.2 Capillary Zone Electrophoresis; 2.3 Electroosmotic Flow; 3 Instrumentation; 3.1 Injection; 3.2 Detection4 Applications; 4.1 Peptide Separations; 4.2 Protein Separations; 5 References; II.4 Microseparation Techniques III: Gel Electrophoresis for Sample Preparation in Protein Chemistry; 1 Introduction; 2 Denaturing Techniques; 2.1Commonly Used SDS-Polyacrylamide Gel Electrophoresis Techniques for Protein Separation; 2.2 Blue-SDS-PAGE for Quantitative Protein Recovery from Gels; 2.3 Electroelution of Proteins After Blue-SDS-PAGE; 2.4 Electroblotting of Blue and Colourless SDS Gels; 2.5 Isoelectric Focusing in the Presence of Urea; 3 Native Techniques; 3.1 Colourless-Native-PAGE; 3.2 Blue-Native-PAGE3.3 Native Isoelectric Focusing; 4 References; II.5 Microseparation Techniques IV: Electroblotting; 1 Introduction; 2 Electroblotting; 2.1 Polyacrylamide Gel Electrophoresis; 2.2 Blot Systems; 2.2.1 Tank Blotting; 2.2.2 Semidry 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; Section III: Amino Acid Analysis; III.1 Amino Acid Analysis; 1 Introduction; 2 Sample Preparation; 2.1 Peptides and Proteins; 2.1.1 Enzymatic Hydrolysis2.1.2 Acid Hydrolysis; 2.1.3 Alkaline Hydrolysis; 2.2 Free Amino Acids; 3 Derivatization; 3.1 Post-Column Derivatization; 3.1.1 Ninhydrin; 3.1.2 Orthophthaldialdehyde; 3.1.3 Fluorescamine; 3.2 Pre-Column Derivatization; 3.2.1 Phenylisothiocyanate; 3.2.2 Orthophthaldialdehyde; 3.2.3 Fluorenylmethyl Chloroformate; 3.2.4 Dabsyl Chloride; 3.2.5 Dansyl Chloride; 3.2.6 Chiral Reagents; 4 Data Evaluation; 5 Instrumentation; 6 Discussion; 7 References; Section IV. Protein Sequence Analysis; IV. 1 The Edman Degradation; 1 The Edman Chemistry; 1.1 Coupling, Cleavage and Conversion
• Sommario/riassunto: Both a thorough introduction and laboratory guide!This book is based on the lectures and demonstrations presented at a successful workshop organized by the authors. Expert contributions and the support of more than a dozen companies engaged in bioanalysis and instrumentation enabled the participants to familiarize themselves with the most recent developments e.g. in protein separation and characterization (including laser desorption-ionisation mass spectrometry), fragmentation and micro sequencing.The workshop was held in the Max Planck Institute for Biochemistry at Martinsried