Titlebook: Lipid Signaling Protocols; Mark G. Waugh Book 2016Latest edition Springer Science+Business Media New York 2016 cell function.ceramides.cho

fatty-acids

Luciferase Reporter Assays to Assess Liver X Receptor Transcriptional Activity,y by transcription factors, and the discovery of gene regulatory elements and small-molecule modulators with high levels of precision. This is made possible through detection of bioluminescence produced by luciferase-coding reporters in a wide range of cellular environments. These assays are routine

somnambulism

Metabolically Biotinylated Reporters for Electron Microscopic Imaging of Cytoplasmic Membrane Microoteins, including ion channels, receptors, and other signaling molecules, exhibit a profound submicroscopic spatial organization, in some cases clustering in submicron membrane subdomains having a protein and lipid composition distinct from that of the bulk membrane. In the case of membrane-associat

Canopy

FLACK

Isolation and Analysis of Detergent-Resistant Membrane Fractions,omains highly enriched in glycosphingolipids, sphingomyelin, ceramide, and cholesterol, was formulated by van Meer and Simons in 1988 and came to a turning point when it was suggested that lipid rafts could be isolated thanks to their resistance to solubilization by some detergents, namely Triton X-

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Detection of Isolated Mitochondria-Associated ER Membranes Using the Sigma-1 Receptor,al functions. A specific marker for this important entity of cellular structure is urgently needed. Thus, we propose in this method chapter that the membrane-bound ER chaperone sigma-1 receptor serves as an ideal marker for the MAM. We describe in detail the preparation and purification of the MAM b

NEX

,Using Surface Plasmon Resonance to Quantitatively Assess Lipid–Protein Interactions,ng a ligand onto a senor chip, measuring a baseline resonance angle, and flowing an analyte in bulk solution over the fixed ligand to measure the subsequent change in resonance angle. The mass of analyte bound to fixed ligand is directly proportional to the resonance angle change and the system is s

刀锋

,Analyzing Protein–Phosphoinositide Interactions with Liposome Flotation Assays,ons more than protein–lipid overlay assays, which makes this method less prone to detect false positive interactions. However, liposome lipid composition must be well-considered in order to prevent nonspecific binding of the protein through electrostatic interactions with negatively charged lipids l

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forager

Analysis of Sphingolipid Synthesis and Transport by Metabolic Labeling of Cultured Cells with [3H]Ss and transporters. The biosynthesis of sphingolipids in cultured cells is initiated in the endoplasmic reticulum (ER) by the formation of a sphingoid base from serine and palmitoyl-CoA. N-acylation of the sphingoid base produces ceramide, which is transported to the Golgi apparatus where phosphocho