Plant Lipid Signaling Protocols 1st Edition by Teun Munnik, Xavier Zarza, Teun Munnik, Ingo Heilmann ISBN 1627034005 9781627034005

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ISBN 10: 1627034005 
ISBN 13: 9781627034005
Author: Teun Munnik, Xavier Zarza, Teun Munnik, Ingo Heilmann

As scientist begin to understand the complexity of lipid signaling and its roles in plant biology, there is an increasing interest in their analysis. Due to the low abundancy and transient nature of some of these hydrophobic compounds, this is not always easy. In Plant Lipid Signaling Protocols, expert researchers in the field detail experimental approaches by which plant signaling lipids can be studied. These methods and techniques include analysis of plant signaling lipids, including detailed protocols to detect various relevant compounds by targeted or non-targeted approaches; to assay relevant enzyme activities in biological material or using recombinant enzymes; to test for specific binding of signaling lipids to protein partners; or to visualize signaling lipids or lipid-derived signals in living plant cells. 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.

Plant Lipid Signaling Protocols 1st Table of contents:

Part I: General Methods and Lipid Extraction

• Chapter 1: Principles of Lipid Extraction from Plant Tissues
  • Overview of Common Lipid Extraction Methods (e.g., Folch, Bligh & Dyer, modified methods)
  • Considerations for Different Plant Tissues and Lipid Classes
  • Minimizing Lipid Degradation and Contamination
• Chapter 2: Lipid Cleanup and Fractionation Techniques
  • Solid-Phase Extraction (SPE) for Lipid Separation
  • Thin-Layer Chromatography (TLC) for Lipid Class Separation
  • Column Chromatography Approaches
• Chapter 3: Lipid Quantification and Derivatization for Analysis
  • Gravimetric Methods
  • Colorimetric Assays (e.g., Phosphate determination)
  • Derivatization for GC-MS and LC-MS Analysis

Part II: Analysis of Specific Lipid Signals and Metabolites

• Chapter 4: Analysis of Phosphoinositides (PIPs) and Their Metabolism
  • Extraction and Separation of PIPs by TLC or HPLC
  • Radioactive Labeling and Immunoprecipitation of PIPs
  • Enzyme Assays for Phosphoinositide Kinases and Phosphatases
  • Imaging PIPs in Live Cells
• Chapter 5: Quantification of Phosphatidic Acid (PA)
  • Methods for PA Extraction and Quantification (e.g., LC-MS/MS)
  • Enzyme Assays for Phospholipase D (PLD) and Phospholipase C (PLC) Activity
  • In vivo Detection of PA Using Biosensors
• Chapter 6: Sphingolipid Analysis in Plants
  • Extraction and LC-MS/MS Analysis of Sphingolipids
  • Enzyme Assays for Sphingolipid Metabolizing Enzymes
  • Analysis of Sphingolipid-Derived Signaling Molecules
• Chapter 7: Oxylipin Profiling and Quantification
  • Extraction and Derivatization of Oxylipins
  • GC-MS and LC-MS/MS Methods for Oxylipin Analysis
  • Analysis of Jasmonates and Related Compounds
• Chapter 8: Fatty Acid Analysis and Lipidomics Approaches
  • Gas Chromatography (GC) for Fatty Acid Composition
  • High-Resolution Mass Spectrometry for Comprehensive Lipid Profiling (Lipidomics)
  • Bioinformatic Tools for Lipidomics Data Analysis

Part III: Studying Lipid-Protein Interactions and Signaling Pathways

• Chapter 9: In Vitro Assays for Lipid-Binding Proteins
  • Lipid Overlay Assays
  • Surface Plasmon Resonance (SPR) for Lipid-Protein Binding
  • Fluorescence-Based Binding Assays
• Chapter 10: In Vivo Approaches to Lipid-Protein Interactions
  • FRET and BRET-Based Biosensors for Lipid Dynamics and Protein Translocation
  • Fluorescence Microscopy for Lipid Localization
  • Co-immunoprecipitation to Detect Protein-Protein Interactions in Lipid Signaling Complexes
• Chapter 11: Genetic and Pharmacological Manipulation of Lipid Signaling
  • Gene Silencing and Overexpression Strategies for Lipid Pathway Enzymes
  • Use of Specific Lipid Signaling Inhibitors and Activators
  • Phenotypic Characterization of Mutants and Drug-Treated Plants

Part IV: Advanced Techniques and Applications

• Chapter 12: Imaging Lipid Signals in Plant Cells and Tissues
  • Confocal Microscopy for Live-Cell Imaging of Lipids
  • Electron Microscopy for Subcellular Lipid Structures
  • Raman Spectroscopy for Label-Free Lipid Imaging
• Chapter 13: Lipid Rafts and Membrane Microdomains
  • Isolation of Lipid Rafts from Plant Membranes
  • Characterization of Lipid and Protein Composition of Rafts
  • Functional Assays for Raft-Mediated Signaling
• Chapter 14: Bioinformatics and Data Analysis in Plant Lipidomics
  • Databases for Plant Lipids
  • Tools for Pathway Analysis and Network Reconstruction
  • Statistical Methods for Large-Scale Lipidomics Data
• Chapter 15: Future Perspectives and Emerging Technologies in Plant Lipid Signaling
  • Advances in Imaging Technologies
  • Integration with Other Omics Data
  • CRISPR-Cas9 for Targeted Lipid Metabolism Engineering

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Tags: Teun Munnik, Xavier Zarza, Teun Munnik, Ingo Heilmann, Plant Lipid, Signaling