Aeris widepore 150 2.1 mm

Ethylene glycol polymers were separated by an Agilent 1100 series LC system (Agilent Technologies, Palo Alto, CA) with an RP-C18 microbore HPLC column (Phenomenex Aeris WIDEPORE 150 2.1 mm, 3- m particle size, 200Å pore diameter). A ThermoFinnigan LCQ Deca-XP mass spectrometer was used to observe ethylene glycol oligomers.

WNK1 autophosphorylation reactions were performed and stopped by the
addition of guanidine to 1 M final concentration at the time points indicated.
WNK1 was digested in an 8:1 molar ratio with sequence-grade chymotrypsin (Roche)
in the presence of 100 mM Tris pH 8.0, and 25 mM CaCl₂ (100
μL total reaction volume) at 30 °C overnight. Following
digestion, the peptide mixture was separated by HPLC (Agilent 1100) on a RP-C18
column (Phenomenex Aeris Widepore 150 × 2.1 mm) using an
acetonitrile-water gradient from 4% to 28% with 0.2%
formic acid. Mass spectrometric analysis was performed on an LCQ DECA XP
ion-trap mass spectrometer (ThermoFinnigan) with the HPLC coupled inline to an
orthogonal electrospray ionization source. MS detector responses were obtained
by integration under ion traces corresponding to m/z ranges for
activation loop peptides, then scaled to the integration of the total ion
current for all eluted peptides. MS/MS spectra were acquired in a data dependent
mode and analyzed using MASCOT software (Matrix Science Ltd.) (71 (link), 72 (link)).
Synthetic peptide standards (21st Century Biochemicals) were used to confirm
HPLC elution times and for quantitation.