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Antigen Processing Methods and Protocols 1st Edition by Peter Van Endert ISBN 1627032177 9781627032179

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Antigen Processing Methods and Protocols 1st Edition Peter Van Endert (Eds.) Digital Instant Download

Author(s): Peter van Endert (eds.)
ISBN(s): 9781627032179, 1627032177
Edition: 1
File Details: PDF, 7.38 MB
Year: 2013
Language: english
SKU: EB-4318092 Category: Tag:

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ISBN 10: 1627032177 
ISBN 13: 9781627032179
Author: Peter Van Endert

Antigen processing is a biological process that prepares antigens for the presentation to special cells in the immune system called T lymphocytes. In Antigen Processing: Methods and Protocols, expert researchers in the field provide a comprehensive set of protocols for studying presentation of antigens produced in the standard processing pathways for MHC class I and class II molecules. The chapters follow chronology of intracellular processing events, ending with recognition of peptide-MHC complexes at the cell surface by T lymphocytes. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Antigen Processing Methods and Protocols 1st Table of contents:

  • Part I: Proteolytic Pathways for MHC Class I Ligand Generation

      1. Purification of 20S Proteasomes from Various Tissues (e.g., Liver, Spleen)
      1. Assays for Proteasome Activity and Peptide Hydrolysis
      1. Analysis of Proteasome-Generated Peptides by Mass Spectrometry
      1. Purification and Characterization of Immunoproteasomes
      1. Studying the Role of Endoplasmic Reticulum Aminopeptidases (ERAPs) in Peptide Trimming
      1. Generating and Analyzing Truncated Peptides in Vitro

  • Part II: Peptide Transport and MHC Class I Assembly

      1. Assays for Transporter Associated with Antigen Processing (TAP) Function (e.g., Peptide Transport Assays, ATP Hydrolysis)
      1. Biochemical Analysis of the Peptide Loading Complex (PLC)
      1. Measuring MHC Class I Surface Expression and Stability
      1. Mutational Analysis of MHC Class I Molecules for Peptide Binding
      1. In Vitro Refolding of Soluble MHC Class I Molecules with Peptides
      1. Analyzing TAP-Independent Antigen Presentation Pathways

  • Part III: Endosomal and Lysosomal Processing for MHC Class II Ligand Generation

      1. Isolation of Antigen-Presenting Cells (APCs) for Class II Studies (e.g., Dendritic Cells, B Cells)
      1. Loading Exogenous Antigens onto MHC Class II Molecules In Vitro
      1. Studying Antigen Uptake and Trafficking in APCs
      1. Biochemical Analysis of Endosomal Compartments Involved in Antigen Processing
      1. In Vitro Assays for Proteolytic Activity in Endosomes (e.g., Cathepsin Assays)
      1. Analysis of Invariant Chain (Ii) Processing and CLIP Removal

  • Part IV: MHC Class II Assembly and Peptide Loading

      1. Investigating the Role of HLA-DM and HLA-DO in Peptide Exchange
      1. Measuring MHC Class II Surface Stability and Half-Life
      1. Analysis of MHC Class II-Peptide Complexes by Mass Spectrometry
      1. Flow Cytometric Analysis of MHC Class II Expression and Conformational Changes

  • Part V: Functional Assays for Antigen Presentation

      1. T-Cell Hybridoma Activation Assays for MHC Class I Presentation
      1. T-Cell Proliferation Assays for MHC Class II Presentation
      1. Cytotoxicity Assays (e.g., Chromium Release Assays for CTLs)
      1. ELISPOT Assays for Cytokine Production by T Cells
      1. Tetramer Staining for Antigen-Specific T Cells
      1. In Vivo Models for Studying Antigen Processing and Presentation

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Tags: Peter Van Endert, Antigen, Methods