NSC-34 cells transfected with vehicle or TDP-43 were washed out in 0.1 M cacodylate buffer (C0250, Sigma-Aldrich) and fixed in 0.1 M cacodylate buffer containing 2.5% glutaraldehyde (16365, Electron Microscopy Sciences) for 1 hour at room temperature. The cells were postfixed in 1% osmium tetroxide (19100, Electron Microscopy Sciences) for 1 hour, 1% tannic acid (W304204, Sigma-Aldrich) for 30 min, and 1% aqueous uranyl acetate (77870.01, SERVA, Heidelberg, DE) for 1 hour. Subsequently, samples were dehydrated through a graded ethanol series and flat-embedded in resin (14120, Electron Microscopy Sciences) for 24 hours at 60°C. Ultrathin sections (50 nm) were cut parallel to the substrate and counterstained with 5% uranyl acetate in 50% ethanol. Electron micrographs were acquired with a Hitachi 7800 120-kV TEM (Hitachi) equipped with MegaView G3 digital camera and RADIUS software (EMSIS) using the Multiple Image Alignment montage tool.
W304204
Manufactured by Merck Group
Sourced in United Kingdom
The W304204 is a laboratory equipment product manufactured by Merck Group. It serves as a general-purpose tool in scientific research and analysis. The core function of this product is to provide a reliable and consistent platform for various laboratory procedures and experiments. No further details or interpretations about its intended use are provided.
Automatically generated - may contain errors
2 protocols using w304204
1
Ultrastructural Analysis of TDP-43 in NSC-34 Cells
2
Evaluating Chemical and Enzymatic Treatments
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Three different chemical compounds and one enzymatic treatment were tested at varying concentrations as preselected from the literature. Solutions of tannic acid (0.05, 0.1 and 0.2 % by weight; C76H52O46; W304204; Sigma-Aldrich, UK), L-Cysteine (1.0 and 2.0 % by weight; C3H7NO2S; W326205; Sigma-Aldrich, UK), sodium sulfite (0.5, 1.0 and 2.0 %; Na2O3S; S0505, Sigma-Aldrich, UK) and the proteolytic enzyme, Alcalase® Bacillus Licheniformis (4.0, 3.0, 2.0, 1.0 and 0.5 %; 126741-500; VWR, UK) were prepared using hatchery water. Sodium sulfite had a measurable effect on salinity (35 and 48 ppt at 0.5 and 2.0 %, respectively) such that each concentration was also prepared and tested in distilled freshwater (1, 3, and 21 ppt at 0.5, 1.0 and 2.0 %, respectively). The pH of each solution was adjusted to that of the hatchery water (pH 8.0) with 5 M HCl or 5 M NaOH using a calibrated pH-meter (Mettler Toledo, MP220/225), and salinity was measured using a handheld refractometer.
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