Mass Spec Parameters: Chromatography run at ~2 uL/min. 11 salt steps were performed on a SCX column. Buffer C run as background. Buffer D run for 10 minutes at: (Percent D): 0, 2, 4, 6, 8, 10, 15, 20, 30, 50, 90. In mM ammonium chloride this is: 000, 020, 040, 060, 080, 100, 150, 200, 300, 500, 900. Then Buffer C was run for 75 minutes. Fractions were run onto a C18 reverse phase column RPC for analysis. RPC chromatography: Minute %BufferA %BufferB 0 95 5 1 95 5 61 55 45 62 5 95 67 5 95 68 95 5 85 95 5 BUFFERS: Buffer A: 99.9% H20, 0.1% Formic Acid Buffer B: 99.9% acetonitrile, 0.1% Formic Acid Buffer C: 95% H20, 5% ACN, 0.1% Formic Acid Buffer D: 95% H20, 5% ACN, 0.1% Formic Acid, 1M ammonium chloride Mass Spectrometer: ThermoFinnigan ESI-Ion Trap "Dexa XP Plus" MS Settings: MS Run Time (min): 85.00 Duration (min): 85.00 Number of Scan Events: 4 Scan Event Details: 1: + c norm o(300.0-1500.0) 2: + c norm Dep MS/MS Most intense ion from (1). 3: + c norm Dep MS/MS 2nd most intense ion from (1). 4: + c norm Dep MS/MS 3rd most intense ion from (1). 2 microscans for MS scan 3 microscans for MS/MS scans. Data Dependent Settings: Parent Mass List: (none) Reject Mass List: (none) Default Charge State: 2 Default Isolation Width: 5.00 Normalized Collision Energy: 35.0 Activation Q: 0.250 Activation Time: 30.000 Min. Signal Required: 10000 Min. MSn Signal Required: 1000 Most intense if no parent masses found not enabled Add/subtract mass not enabled Charge state screening not enabled Charge state rejection not enabled Global Data Dependent Settings: Exclusion mass width by mass Exclusion mass width low: 1.50 Exclusion mass width high: 1.50 Reject mass width by mass Reject mass width low: 0.50 Reject mass width high: 0.50 ZoomScan mass width by mass ZoomScan mass width low: 5.00 ZoomScan mass width high: 5.00 MS mass range: 0.00-1000000.00 MSn mass range by mass MSn mass range: 0.00-1000000.00 Analog UV data dep. not enabled Dynamic exclusion enabled Repeat Count: 2 Repeat Duration: 0.50 Exclusion List Size: 50 Exclusion Duration: 3.00 Exclusion mass width by mass Exclusion mass width low: 1.50 Exclusion mass width high: 1.50 Isotopic data dependence not enabled Mass Tags data dependence not enabled Custom Data Dependent Settings: Not enabled