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Thisvolume encompasses all major methodologies to interrogate endocannabinoidsystems (ECS) and endocannabinoids (eCBs) signaling. With increasing interesttowards the manifold activities of eCBs, this book discusses the chemical,biochemical, and molecular biological assays, and activity of distinct elementsof the ECS. These include membrane, nuclear receptors, biosynthetic andhydrolytic enzymes, and membrane transporters and oxidative enzymes. Written inthe highly successful Methods in Molecular Biology series format,chapters include introductions to their respective topics, lists of thenecessary materials and reagents, step-by-step, readily reproducible laboratoryprotocols, and tips on troubleshooting and avoiding known pitfalls.
Timely and cutting edge, Endocannabinoid Signaling: Methods and Protocolsis a valuable resource and will help chemists, drug designers, biochemists,molecular biologists, cell biologists, pharmacologists, and (electro)physiologists navigate the mare magnum of endocannabinoid research.
List of contents
Need for Methods to InvestigateEndocannabinoid Signaling.- Extraction and Simultaneous Quantificationof Endocannabinoids and Endocannabinoid-Like Lipidsin Biological Tissues.- Determination of 2-Arachidonoylglycerol by µSPE-LC-MS/MS.- Analysis of Omega-3 Fatty AcidDerived N-acylethanolamines in BiologicalMatrices.- Assay of CB1 Receptor Binding.- The Displacement Binding Assay Using HumanCannabinoid CB2 Receptor-Transfected Cells.- Assay of TRPV1 Receptor Signaling.- A Functional Assay for GPR55: Envision Protocols.- The Cyclic AMP Assay Using Human CannabinoidCB2 Receptor-Transfected Cells.- Assay of GTPGammaS Binding in Autoradiography.- Protocol to Study beta-Arrestin Recruitment by CB1 and CB2Cannabinoid Receptors.- Assay of NAT Activity.- Assay of NAPE-PLD Activity.- Assay of FAAH Activity.- Assay of NAAA Activity.- Assay of DAGL /beta Activity.- Assay of Monoacylglycerol Lipase Activity.- A Sensitive and Versatile Fluorescent Activity Assay for ABHD6.- A Sensitive and Versatile Fluorescent Activity Assay for ABHD12.- Assay of Endocannabinoid Uptake.- Assay of Endocannabinoid Oxidation by Cyclooxygenase-2.- Oxygenation of Anandamide by Lipoxygenases.- Assay of Endocannabinoid Oxidation by Cytochrome P450.- Assessing Gene Expression of the Endocannabinoid System.- Western Blotting of the Endocannabinoid System.- Quantitation of Plasma Membrane (G Protein-Coupled) Receptor Traffickingin Cultured Cells.- Measuring ECS Interaction with Biomembranes.- Visualization of Endocannabinoids in the Cell.
Summary
This
volume encompasses all major methodologies to interrogate endocannabinoid
systems (ECS) and endocannabinoids (eCBs) signaling. With increasing interest
towards the manifold activities of eCBs, this book discusses the chemical,
biochemical, and molecular biological assays, and activity of distinct elements
of the ECS. These include membrane, nuclear receptors, biosynthetic and
hydrolytic enzymes, and membrane transporters and oxidative enzymes. 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 tips on troubleshooting and avoiding known pitfalls.
Timely and cutting edge, Endocannabinoid Signaling: Methods and Protocols
is a valuable resource and will help chemists, drug designers, biochemists,
molecular biologists, cell biologists, pharmacologists, and (electro)
physiologists navigate the mare magnum of endocannabinoid research.
