Download Endocannabinoid Signaling: Methods and Protocols by Mauro Maccarrone PDF

By Mauro Maccarrone

This quantity encompasses all significant methodologies to interrogate endocannabinoid platforms (ECS) and endocannabinoids (eCBs) signaling. With expanding curiosity in the direction of the manifold actions of eCBs, this e-book discusses the chemical, biochemical, and molecular organic assays, and task of special components of the ECS. those comprise membrane, nuclear receptors, biosynthetic and hydrolytic enzymes, and membrane transporters and oxidative enzymes. Written within the hugely winning Methods in Molecular Biology series layout, chapters contain introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, conveniently reproducible laboratory protocols, and pointers on troubleshooting and heading off recognized pitfalls.

Timely and leading edge, Endocannabinoid Signaling: tools and Protocols is a precious source and may aid chemists, drug designers, biochemists, molecular biologists, telephone biologists, pharmacologists, and (electro) physiologists navigate the mare magnum of endocannabinoid research.

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1 Specimens Collection Tissues must be snap-frozen in liquid nitrogen after collection (see Note 1) and stored at −80 °C (see Note 2). 2 Plasma 1. Collect blood in a Vacutainer tube with sodium citrate. 2. Centrifuge blood in the collection tube for 15 min at 150 × g with brake off switch. 3. Remove the tube from the centrifuge. 4. Transfer plasma (top layer) to a 15 ml centrifuge tube. Be careful not to aspirate cells from the buffy coat (cellular) layer. 5. Centrifuge plasma in the 15 ml centrifuge tube for 15 min at 1000 × g.

Biochim Biophys Acta 1811:706–723 2. Buczynski MW, Parsons LH (2010) Quantification of brain endocannabinoid levels: methods, interpretations and pitfalls. Br J Pharmacol 160: 423–442 3. Lerner R, Lutz B, Bindila L (2013) Tricks and tracks in the identification and qualification of endocannabinoids. eLS. Wiley, Chichester. a0023407 4. Duerr GD, Heinemann JC, Gestrich C et al (2014) Impaired border zone formation and adverse remodeling after reperfused myocardial infarction in cannabinoid CB2 receptor deficient mice.

4. 0 ml plasma. The ACN will precipitate all proteins and highly lipophilic compounds, while keeping the endocannabinoids in solution. The ACN will completely mix with the plasma. While adding this solution to the plasma, keep the tube on a vortexer running at a gentle pace. Vortexing the sample will improve the reproducibility of the precipitation. 5. Centrifuge the 15 ml tubes at 3000 × g for 5 min at room temperature. 6. After centrifugation, collect the supernatant and transfer to a clean 50 ml tube.

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