id	author	title	date	pages	extension	mime	words	sentence	flesch	summary	cache	txt
fnp-2749	Oullier, Thibauld; Prigent, Alice; Chapelet, Guillaume; Neunlist, Michel; Letournel, Franck; Derkinderen, Pascal	Optimizing filter trap assay for the detection of aggregated alpha-synuclein in brain samples	2020		.htm	application/xhtml+xml	1678	80	39	Brain samples were lysed with the “Precellys 24™” tissue homogenizer (Bertin technologies, Cat# P000669-PR240-A, Montigny-le-Bretonneux, France) followed by a brief sonication (one minute, 5 seconds on, 5 seconds off, amplitude 70%; VCX130 ultrasonic processor with a 6 mm probe, Sonics and Materials, Newton, CT, USA) either in (i) radioimmunoprecipitation assay (RIPA) buffer (Merck Millipore, Cat# 20-188, Molsheim, France) containing 2 mM orthovanadate (Sigma, Cat# S6505, Saint Quentin Fallavier, France), 1% (v/v) phosphatase inhibitor cocktail III (Sigma, Cat# P0044) and a protease inhibitors cocktail (Roche, Cat# 14424700, Meylan, France) at 4°C; (ii) ‘Sodium dodecyl sulfate (SDS) buffer’ containing 1% (w/v) SDS, 2 mM orthovanadate (Sigma, Cat# S6505) and 1% (v/v) phosphatase inhibitor cocktail III (Sigma, Cat# P0044); (iii) ‘Urea buffer’ containing 7 M urea (Sigma, Cat# T7875), 2 M thiourea (Sigma, Cat# L3771), 4% (v/v) CHAPS (Sigma, Cat# C3023), 2 mM orthovanadate (Sigma, Cat# S6505), 1% (v/v) phosphatase inhibitor cocktail III (Sigma, Cat# P0044) and a protease inhibitors cocktail (Roche, Cat# 14424700). Lysates were centrifuged at 10,000 x g for 5 minutes; supernatants were recovered and protein concentration determined using the Pierce BCA Protein Assay Kit (ThermoFisher Scientific, Cat# 23227, Saint Herblain, France) along with the Varioskan™ multimode microplate reader (ThermoFisher Scientific, Cat# VL0L00D0); samples lysed in urea buffer were diluted 5 times for compatibility with the BCA assay (https://assets.thermofisher.com/TFS-Assets/LSG/Application-Notes/TR0068-Protein-assay-compatibility.pdf).	cache/fnp-2749.htm	txt/fnp-2749.txt