UID:
almahu_9948026500102882
Umfang:
1 online resource (369 p.)
ISBN:
1-281-03314-6
,
9786611033149
,
0-08-053870-3
Inhalt:
Both novices and experts will benefit from this insightful step-by-step discussion of phage display protocols.Phage Display of Peptides and Proteins: A Laboratory Manual reviews the literature and outlines the strategies for maximizing the successful application of phage display technology to one's research. It contains the most up-to-date protocols for preparing peptide affinity reagents, monclonal antibodies, and evolved proteins.Key Features* Prepared by experts in the field* Provides proven laboratory protocols, troubleshooting, and tips* Includes maps, seque
Anmerkung:
Description based upon print version of record.
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Front Cover; Phage Display of Peptides and Proteins: A Laboratory Manual; Copyright Page; Contents; Contributors; Foreword; Preface; Chapter 1. Biology of the Filamentous Bacteriophage; Introduction to the Life Cycle of the Bacteriophage; The Genome and Its Products; The Ff Bacteriophage Particle; The Infection Process; The Assembly Process; References; Chapter 2. Principles and Applications of Phage Display; Introduction; Phage Display of Natural Peptides; Phage Display of Random Peptides; Phage Display of Protein and Protein Domains; Conclusion; World Wide Web Page; References
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Chapter 3. Vectors for Phage DisplayPhagemid; Markers; Other Viral Display Vectors; Proteolysis; Valency; Issue of Biological Selection; References; Chapter 4. Microbiological Methods; Protocol 1: Spreading Techniques; Protocol 2: Titering Bacteriophage; Protocol 3: Preparing Phage Particles; Protocol 4: Preparation of Single-Stranded DNA from Viral Particles; Protocol 5: Preparation of Vector RF DNA; References; Chapter 5. Construction of Random Peptide Libraries in Bacteriophage MI3; Introduction; Protocol 1: Assembly of Double-Stranded DNA Insert from Degenerate Oligonucleotides
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Protocol 2: Preparation of Linearized Vector DNAProtocol 3: Ligation of the Vector and Insert DNA; Protocol 4: Electroporation of Ligated DNA into Bacteria; Protocol 5: Amplification and Harvest of Library Phage; References; Chapter 6. Construction and Screening of Antibody Display Libraries; General Introduction and Background; Production of Antibody Genes and Preparation of Libraries; Protocol 1: Preparation of Lymphocytes as Single-Cell Suspensions from Mouse Spleen Cells; Protocol 2: Preparation of Human Peripheral Blood Lymphocytes
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Protocol 3: Preparation of Mouse or Human Antibody cDNAConstruction of Antibody V Gene Repertoire; Protocol 4: Preparation and Purification of Primary VH and VL PCR Products; Protocol 5: Preparation and Purification of Linker Fragments; Protocol 6: Assembly of Human Single-Chain Fv Antibody Fragments; Protocol 7: Restriction Digestion of Assembled PCR Fragments; Protocol 8: Preparation of Vector DNA; Protocol 9: Ligation of Insert into Vector; Manipulation of Libraries; Protocol 10: Rescuing Phagemid Libraries; Protocol 11: Selection of Phage-Antibody Libraries by Panning in ""Immunotubes""
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Protocol 12: Selection of Phage-Antibody Libraries by Biotin SelectionProtocol 13: Introducing Diversity by Chain Shuffling; Screening and Expression of Selected Clones; Protocol 14: Induction of Soluble Antibody Fragments from Phagemid Vectors; Protocol 15: Preparation of Periplasmic Fraction and Purification of Antibodies by Immobilized Metal Affinity Chromatography (IMAC); Protocol 16: Small-Scale Rescue of Phagemid Particles Using Helper Virus; Protocol 17: ELISA Using either Soluble Fragments or Phage Particles; References; Chapter 7. Phagemid-Displayed Peptide Libraries; Introduction
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Materials
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English
Weitere Ausg.:
ISBN 0-12-402380-0
Sprache:
Englisch
