Proteomic Profiling

1. Front Matter

   Pages i-xv

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2. How Modern Mass Spectrometry Can Solve Ancient Questions: A Multi-Omics Study of the Stomach Content of the Oldest Human Ice Mummy, the 5300-Year-Old Iceman or Oetzi

   • Rudolf Grimm

   Pages 1-12

3. Step-by-Step Sample Preparation of Proteins for Mass Spectrometric Analysis

   • Xinping Li, Thomas Franz, Ilian Atanassov, Thomas Colby

   Pages 13-23

4. Lab-on-a-Filter Techniques for Economical, Effective, and Flexible Proteome Analysis

   • Yi-Han Lin, Rodrigo Vargas Eguez, Isha Vashee, Yanbao Yu

   Pages 25-34

5. Released N-Glycan Analysis for Biotherapeutic Development Using Liquid Chromatography and Mass Spectrometry

   • Ximo Zhang

   Pages 35-53

6. Comprehensive Protocol to Simultaneously Study Protein Phosphorylation, Acetylation, and N-Linked Sialylated Glycosylation

   • Marcella Nunes Melo-Braga, María Ibáñez-Vea, Katarzyna Kulej, Martin R. Larsen

   Pages 55-72

7. Phos-Tag Fluorescent Gel Staining for the Quantitative Detection of His- and Asp-Phosphorylated Proteins

   • Emiko Kinoshita-Kikuta, Eiji Kinoshita, Tohru Koike

   Pages 73-78

8. Polyubiquitin Profile in Down Syndrome and Alzheimer’s Disease Brain

   • Antonella Tramutola, Marzia Perluigi

   Pages 79-91

9. Platform Methods to Characterize the Charge Heterogeneity of Three Common Protein Therapeutics by Imaged Capillary Isoelectric Focusing

   • Jiaqi Wu, Will McElroy, Charles Haitjema, Carsten Lück, Christopher Heger

   Pages 93-103

10. A Protocol for Isolation, Purification, Characterization, and Functional Dissection of Exosomes

    • Alin Rai, Haoyun Fang, Monique Fatmous, Bethany Claridge, Qi Hui Poh, Richard J. Simpson et al.

    Pages 105-149

11. Human Plasma Extracellular Vesicle Isolation and Proteomic Characterization for the Optimization of Liquid Biopsy in Multiple Myeloma

    • Antonia Reale, Tiffany Khong, Rong Xu, Maoshan Chen, Sridurga Mithraprabhu, Nicholas Bingham et al.

    Pages 151-191

12. Isolation of Extracellular Vesicles for Proteomic Profiling

    • Dongsic Choi, Janusz Rak, Yong Song Gho

    Pages 193-206

13. Isolation of Proteins from Extracellular Vesicles (EVs) for Mass Spectrometry-Based Proteomic Analyses

    • Prabal Subedi, Michael Schneider, Michael J. Atkinson, Soile Tapio

    Pages 207-212

14. Flow Cytometry as an Important Tool in Proteomic Profiling

    • Michael P. Blundell, Sharon L. Sanderson, Tracey A. Long

    Pages 213-227

15. Improved Immunoprecipitation to Mass Spectrometry Method for the Enrichment of Low-Abundant Protein Targets

    • Penny Jensen, Bhavin Patel, Suzanne Smith, Renuka Sabnis, Barbara Kaboord

    Pages 229-246

16. Multiplex Fluorescent Bead-Based Immunoassay for the Detection of Cytokines, Chemokines, and Growth Factors

    • Jonathan M. Preuss, Ute Burret, Sabine Vettorazzi

    Pages 247-262

17. Bead-Based Multiplex Immunoassays: Procedures, Tips, and Tricks

    • Candice Cox

    Pages 263-276

18. Immunoaffinity-Based Liquid Chromatography Mass Spectrometric Assay to Accurately Quantify the Protein Concentration of HMGB1 in EDTA Plasma

    • Viktoria Anselm, Andreas Steinhilber, Cornelia Sommersdorf, Oliver Poetz

    Pages 277-289

19. Recombinant Anti-idiotypic Antibodies in Ligand Binding Assays for Antibody Drug Development

    • Stefan Harth, Christian Frisch

    Pages 291-306

20. cDNA Display-Mediated Immuno-PCR (cD-IPCR): An Ultrasensitive Immunoassay for Biomolecular Detection

    • Chathuni Jayathilake, Naoto Nemoto

    Pages 307-321

This detailed new edition presents the latest developments of the main pillars of protein analysis, namely sample preparation, separation, and characterization. Core areas in this volume are protocols for the analysis of post-translational modifications and protein interaction partners, followed by sophisticated procedures to enrich for extracellular vesicles and organelles, along with several types of protein immuno-assays complemented by various methods for the characterization of antibodies and host-cell protein analysis. Last but not least, a few standard sample preparation protocols and recent advances concerning immuno-chemical detection of proteins are included as well. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and up-to-date, Proteomic Profiling: Methods and Protocols, Second Edition serves as an ideal reference for students of biochemistry, biomedicine, biology, and genomics and will be an invaluable source for the experienced, practicing scientist as well.

• Protein interaction partners
• Mass spectrometric analysis
• Protein immuno-assays
• Extracellular vesicles and organelles
• Immuno-chemical detection
• Protein analysis

• Bio-Rad Laboratories GmbH, Feldkirchen, Germany

  Anton Posch

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