Optima lc ms grade acn

Isotopic reagents for label synthesis including leucines (L-leucine, L-leucine-1-¹³C, ¹⁵N, and L-leucine-1-¹³C), heavy formaldehydes (CD₂O and ¹³CD₂O), sodium cyanoborodeuteride (NaBD₃CN), 97% ¹⁸O water (H₂¹⁸O) and deuterium water (D₂O) were obtained from ISOTEC Inc. (Miamisburg, OH). Sodium cyanoborohydride (NaBH₃CN), formaldehyde (CH₂O), hydrogen chloride gas (HCl), iodoacetamide (IAA), tris hydrochloride, reagent-grade formic acid (FA), triethylammonium bicarbonate (TEAB), N,N-dimethylformamide (DMF), 4-(4, 6-dimethoxy-1, 3, 5-triazin-2-yl)-4-methylmorpholinium tetrafluoroborate (DMTMM), N-methylmorpholine (NMM), and bovine serum albumin (BSA) were purchased from Sigma-Aldrich (St. Louis, MO). Neuropeptide standard allatostatin I (AST-I, GDGRLYAFGL-NH₂) was purchased from American Peptide Company (Sunnyvale, CA). Sequencing grade trypsin was acquired from Promega (Madison, WI). Urea, ammonium bicarbonate, ACS grade methanol (MeOH), dichloromethane (DCM, CH₂Cl₂), and acetonitrile (ACN, C₂H₃N) were purchased along with Optima LC/MS grade ACN, water, and formic acid from Fisher Scientific (Pittsburgh, PA). The 8-plex iTRAQ reagents were provided by AB SCIEX (Foster City, CA) while TMT⁰ was provided by Thermo Pierce (Rockford, IL).

Chemicals were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA), unless stated otherwise. Carbo-free blocking solution was purchased from Vector Laboratory (Burlingame, CA, USA). SuperSignal West Pico Chemiluminescent Substrate was purchased from Thermo Scientific (Rockford, IL, USA). Precast gel (4–20%) and Precision Plus Protein Standard Kaleidoscope Pre-stained protein markers were purchased from Bio-Rad (Hercules, CA, USA). Percoll was purchased from Pharmacia (Piscataway, NJ, USA). NP-40 was purchased from Calbiochem EMD Millipore (Billerica, MA, USA). SimplyBlue SafeStain was purchased from Life Technologies (Carlsbad, CA, USA). The BCA assay kit was purchased from Thermo Scientific. Biotinylated glycans were obtained from Lectinity (Moscow, Russia). ACS grade methanol (MeOH), ACS grade acetonitrile (ACN), Optima LC-MS grade ACN, Optima LC-MS grade water and ammonium bicarbonate were purchased from Fisher Scientific (Pittsburgh, PA, USA). Deionized water (18.2 MΩ cm) was prepared with a Milli-Q Millipore system. Formic acid (≥98%) was obtained from Fluka (Buchs, Switzerland). DL-Dithiothreitol (DTT), Protease-Max surfactant and sequencing-grade modified trypsin were purchased from Promega (Madison, WI, USA). Recombinant lactadherin protein was purchased from R&D Systems (Minneapolis, MN, USA; product 2805-MF/CF).