8 m guanidine hydrochloride guhcl

Ammonium bicarbonate (ABC), Dowex cation-exchange resin (50W-X8), trifluoroacetic acid (TFA), hydrochloric acid (HCl), DL-dithiothreitol (DTT), ammonium acetate, sodium chloride (NaCl) and sodium borohydride (NaBH4) were obtained from Sigma-Aldrich (Steinheim, Germany). The 8 M guanidine hydrochloride (GuHCl) was purchased from Thermo Fisher Scientific. Peptide N-glycosidase F (PNGase F) was purchased from Roche Diagnostics (Mannheim, Germany). Glacial acetic acid and potassium hydroxide (KOH) were purchased from Honeywell Fluka. Solid phase extraction (SPE) bulk sorbent Carbograph was obtained from Grace Discovery Sciences (Columbia, MD, USA). HPLC SupraGradient acetonitrile (ACN) was obtained from Biosolve (Valkenswaard, The Netherlands) and other reagents and solvents such as methanol, ethanol and 2-propanol were purchased from Merck (Darmstadt, Germany). MultiScreen^® HTS 96-multiwell plates (pore size 0.45 μm) with high protein-binding membranes (hydrophobic Immobilon-P PVDF membrane) and a 96-well PP Microplate were purchased from Millipore (Amsterdam, The Netherlands), conical 96-well Nunc plates from Thermo Scientific (Roskilde, Denmark) and a 96-well PP filter plate from Orochem Technologies (Naperville, IL, USA). Ultrapure water was used for the all preparations and washes, generated from a Q-Gard 2 system (Millipore, Burlington, MA, USA).