Synergy uv system

Sucrose, fluorescein, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), mannitol, ethaminetetraacetic acid (EDTA), poly-(vinyl alcohol) (99%+ hydrated, molecular weight 89,000–980,000), dimethyl sulfoxide (DMSO), and ethylene-bis-(oxyethylenenitrilo)tetraacetic acid (EGTA) were obtained from Sigma-Aldrich (St. Louis, MO). Tris base, 4-morpholine-propanesulfonic acid (MOPs), sodium chloride, 10 000 units penicillin, 10 mg streptomycin/mL, Dulbecco’s modified Eagle medium (DMEM) high glucose solution, fetal bovine serum, 0.5% trypsin-EDTA (10× solution), AlignFlow flow cytometry beads (2.5 µm), and 200 mM l-glutamine solution (1000×) were obtained from Thermo Fisher Scientific (Waltham, MA). Phosphate buffered saline (PBS, 10× concentration, 1.37 M NaCl, 27 mM KCl, 80 mM Na₂HPO₄, and 20 mM KH₂PO₄, pH 7.4) was obtained from Bio-Rad (Hercules, CA). A polyclonal Dylight488 conjugated rabbit anti-LC3 antibody, was obtained from Novus Biologicals (Littleton, CO). Dylight-488 conjugated rabbit isotype control was obtained from Abcam (Cambridge, UK). Water was purified with a Millipore Synergy UV system (18.2mΩ/cm, Bedford, MA). Cell homogenization buffer consisted of 70 mM Sucrose, 215 mM mannitol, 4.31 mM HEPES, and 4.94 mM EDTA, pH 7.4. CE buffer consisted of 250 mM Sucrose, 10 mM HEPES, pH 7.4. PBS was diluted 1:10 in purified water.

Sodium phosphate and sodium citrate, LC-MS grade water, acetonitrile, formic acid and hydrogen peroxide were purchased from Fisher Chemicals (Fair Lawn, NJ). Adenine and L-glutamine were purchased from Acros Biologicals. Catalase was purchased from Sigma (St. Louis, MO) Methionine amide was obtained from Bachem (Torrance, CA). Sequencing-grade modified trypsin was purchased from Promega Corp. (Madison, WI). All reagents were used without further purification. Purified water (18.2 MΩ) was obtained from a Synergy UV system (Millipore, Billerica, MA). Adalimumab was obtained from GlycoScientific (Athens, GA).