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2 protocols using 1.0 m naoh

1

Glucose Uptake in L6 Muscle Cells

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KCl was obtained from ACP Inc (Montreal, QC, Canada) and Na2SO4 from Laboratoire MAT (Québec city, QC, Canada). Formic acid, 1.0 M HCl, and 1.0 M NaOH solutions were from Fisher Scientific (Montreal, QC, Canada), trifluoroacetic acid was purchased from J.T. Baker (Phillipsburg, NJ, USA). NaCl, Acetonitrile optima® liquid chromatography-mass spectrometry (LC/MS), and water grade were from VWR international (Montréal, QC, Canada). Concerning the glucose uptake experiments, the alpha-Minimal Essential Medium (α-MEM), Fetal Bovine Serum (FBS), and trypsin (0,25% solution) were obtained from Invitrogen (Burlington, ON, Canada). The 2-déoxy-D-glucose (non-radioactive), CaCl2, Hepes-Na, and MgSO4 were purchased from Sigma Aldrich (Oakville, ON, Canada). D-2-deoxy-[3H] glucose was from Perkin Elmer (Woodbridge, ON, Canada) and Pierce® BCA Protein Assay Kit BCA was from Pierce Biotechnology (Rockford, IL, USA). Insulin was from CHUL’s pharmacy (Québec, QC, Canada). Also, the L6 skeletal muscle cells line, derived from neonatal rat thigh skeletal muscle, were provided by Dr. A. Klip, Hospital for Sick Children (Toronto, ON, Canada).
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2

Synthesis and Characterization of Fe(OH)2

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Fe(OH) 2 (s) Synthesis. A detailed synthesis procedure for Fe(OH) 2 (s) and product characterization may be found in previously published work. 18 Briefly, Fe(OH) 2 (s) was prepared and stored inside an anoxic chamber (Coy Laboratories) that was maintained using a gas mix of N 2 (98%) and H 2 (2%). Fe(II)Cl 2 •4H 2 O (14 g, >95%, Fisher Scientific) was dissolved in N 2 -purged double deionized water (400 g, DDI, Millipore 18Ω). Dissolved Fe(II) was then precipitated as Fe(OH) 2 (s) by adding 8.2 mL of 10 M NaOH (Fisher Scientific) to solution and mixing by hand. The solid was allowed to react overnight and then separated from the supernatant using a 0.45 µm Nalgene® filter. The Fe(OH) 2 (s) was then allowed to dry for 24 hours before it was powdered using a mortar and pestle. Simulant Preparation. A 5 M Na Hanford LAW simulant with 1080 ppm Cr(VI) was generated as described previously, 17 and spiked with 1-100 ppm Tc(VII), using a 10,000 mg/L Tc stock solution (NH 4 TcO 4 ). The starting simulant was characterized using ion chromatography (IC) and inductively coupled plasma optical emission spectrometry (ICP-OES) (Table 1).
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