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2 protocols using 1.0 m hcl

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

Surface-Functionalized Polystyrene Microbeads

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Polystyrene microbeads functionalized with surface carboxyl groups (diameter = 0.99 μm (μP1), 1.04 μm (μP2), and 0.20 μm (μP3)) were obtained from Bangs Laboratories, Inc. (Fishers, IN); FC04N, PC04N, and PC02N, respectively. BODIPY disulfonate fluorophore (BODIPY2–) was obtained from Molecular Probes (Eugene, OR). Pluronic F108 was purchased from BASF (Florham Park, NJ). Tris–HCl buffer solution was prepared by dissolving reagent grade Trizma base (Sigma-Aldrich, St. Louis, MO) in deionized water (18.0 MΩ cm, Milli-Q Gradient System, MilliporeSigma, Burlington, MA) and then adjusting the solution pH to 8.1 using 1.0 M HCl (Fisher Scientific, Hampton, NH). Microfluidic channels were fabricated using poly(dimethylsiloxane) (PDMS) prepared from a silicone elastomer kit (Sylgard 184, Dow, Midland, MI).
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