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Proteins, Expression, Isolation and Analysis | ... identification. Positive signals were visualized with ONE-HOUR Western™ Detection System (Genscript) using 3,3,5,5-Tetramethylbenzidine (Sigma) as substrate. Negative Stain Electron Microscopy and class-averaging. In-house ... | Get A Quote |
An important step in the production of inflammatory mediators of the leukotriene family is the Ca2+ mediated recruitment of 5 Lipoxygenase (5LO) to nuclear membranes. To study this reaction in vitro, the natural membrane mimicking environment of nanodiscs was used. Nanodiscs with 10.5 nm inner diameter were made with the lipid POPC and membrane scaffolding protein MSP1E3D1. Monomeric and dimeric 5LO were investigated. Monomeric 5LO mixed with Ca2+ and nanodiscs are shown to form stable complexes that 1) produce the expected leukotriene products from arachidonic acid and 2) can be, for the first time, visualised by native gel electrophoresis and negative stain transmission electron microscopy and 3) show a highe... More