Leupeptin

The following materials were from Sigma-Aldrich (St. Louis, MO): guanidine hydrochloride (GdmCl), S-methylmethanesulfonate (MMTS), dimethyl sulfoxide (DMSO), urea, centrifugal filter units (Amicon Ultra-0.5, 10k), tris(hydroxymethyl)aminomethane hydrochloride (Tris•HCl), thermolysin from geobacillus stearothermophilus, proteinase K from tritirachium album, phenylmethylsulfonyl fluoride (PMSF), trifluoroacetic acid (TFA), triethylammonium bicarbonate buffer (TEAB, 1 M, pH 8.5), methacrylic acid N-hydroxysuccinimide(NHS) ester, cytochrome c from equine heart. The following materials were from Thermo-Scientific (Waltham, MA): acetonitrile (ACN, LC-MS grade), 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF), Bestatin, E-64, Leupeptin, Pepstatin A, EDTA solution (Corning, 0.5M, pH 8.0), TMT10plex isobaric label reagent set, N-hydroxysuccinimide(NHS)-activated agarose dry resin (Pierce), trypsin protease MS grade (Pierce). Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) was from Santa Cruz Biotechnology (Dallas, TX). Phosphate-buffered saline (PBS, pH 7.4, 1X) was from Gibco (Gaithersburg, MD). Geldanamycin (≥98 wt%) was from Chem-Impex International, Inc. (Wood Dale, IL). Cyclosporin A (CsA) was from LKT Laboratories, Inc. MacroSpin columns (silica C4 and C18) were from Nest Group (Southborough, MA).

The following materials were from Sigma-Aldrich (St. Louis,MO): guanidine hydrochloride (GdmCl), S-methylmethanethiosulfonate (MMTS), dimethyl sulfoxide (DMSO), urea, centrifugal filter units (Amicon Ultra-0.5 mL, 10 K MWCO), tris(hydroxymethyl)aminomethane hydrochloride (Tris-HCl), thermolysin from geobacillus stearothermophilus, trifluoroacetic acid (TFA), triethylammonium bicarbonate buffer (TEAB, 1 M, pH 8.5), hydrogen peroxide (H₂O₂) (30% w/w), acetic acid, and 2-mercaptoethanol. The following materials were from ThermoScientific (Waltham, MA): acetonitrile (ACN, LC-MS grade), 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF), bestatin, E-64, leupeptin, pepstatin A, EDTA solution (Corning, 0.5M, pH 8.0), TMT10-Plex isobaric label reagent set, NHS-activated agarose dry resin (Pierce), and porcine pancreas trypsin (proteomics grade). Methanol (gradient grade OmniSolv®) was from EMD Millipore. Both Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) and Cyclosporine A (CsA) were from Santa Cruz Biotechnology (Dallas, TX). Phosphate-buffered saline (PBS, pH 7.4) was from Gibco (Gaithersburg, MD). MacroSpin columns (silica C18) and Pi³™ Methionine reagent kit were from Nest Group (Southborough, MA).