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E-raamat: Phosphoinositides: Methods and Protocols

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Phosphoinositides: Methods and ProtocolsSuurem pilt
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This detailed book examines experimental approaches used to investigate the regulation and function of phosphoinositides (PtdInsP), rare eukaryotic phospholipids with a broad role in biological processes such as signal transduction, cell migration and adhesion, cell growth, subcellular organization, and membrane trafficking. The combination of complementary biochemical, mass spectrometry, and imaging methods are instrumental for the detection and quantification of PtdInsP species, as well as induced dimerization methods, affinity precipitation or co-sedimentation with liposomes, protein insertion within lipid bilayers, and enzymatic assays, or through emerging methods like native mass spectrometry and microfluidics, all of which are touched upon in this volume. Written in the highly successful Methods in Molecular Biology format, 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 practical, Phosphoinositides: Methods and Protocols provides detailed methodology for both specialist and novice researchers on a variety of complementary methods that have been instrumental in dissecting the regulation, dynamics, and function of PtdInsPs.
Simultaneous Detection of Phosphoinositide Lipids by Radioactive
Metabolic Labeling.- Label-Free Quantification of Phosphoinositides in
Drosophila by Mass Spectrometry.- Profiling of Phosphoinositide Molecular
Species in Resting or Activated Human or Mouse Platelets by a LC-MS Method.-
Quantification of Genetically-Encoded Lipid Biosensors.- Detection of Plasma
Membrane Phosphoinositide Dynamics Using Genetically-Encoded Fluorescent
Protein Probes.- Imaging the Nanoscale Distribution of Phosphoinositides in
the Cell Plasma Membrane with Single-Molecule Localization Super-Resolution
Microscopy.- Induced Dimerization Tools to Deplete Specific
Phosphatidylinositol Phosphates.- A Real-Time Phosphatidylinositol
4-Phosphate 5-Kinase Assay Using Fluorescence Spectroscopy.- Assessing In
Situ Phosphoinositide-Protein Interactions through Fluorescence Proximity
Ligation Assay in Cultured Cells.- Characterization of
Protein-Phospholipid/Membrane Interactions Using a Membrane-on-a-Chip
Microfluidic System.- Exploring Phosphoinositide Binding Using Native Mass
Spectrometry.- Liposome-Based Methods to Study Protein-Phosphoinositide
Interaction.- Liposome-Based Methods to Study GTPases Activation by
Phosphoinositides.- Liposome Co-Sedimentation and Co-Flotation Assays to
Study Lipid-Protein Interactions.- Characterization of
PROPPIN-Phosphoinositide Binding by Stopped-Flow Fluorescence Spectroscopy.-
Fluorescence Assays to Study Membrane Penetration of Proteins.- Fluorogenic
XY-69 in Lipid Vesicles for Measuring Activity of Phospholipase C Isozymes.
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