5. Acquity UPLC M-Class Symmetry C18 Trap Column, 2D
(100 A ˚ , 5 μm, 180 μm  20 mm) (Waters Corporation).
6. Acquity UPLC M-Class HSS T3 Column (1.8 μm,
75 μm  150 mm) (Waters Corporation).
7. NanoLock Spray dual electrospray ion source (Waters
Corporation).
8. 100 fmol/μL human [Glu
1 ]-fibrinopeptide B human
(Glu-Fib).
9. Solvent C: 20 mM ammonium formiate (pH 10).
10. Solvent D: 100% acetonitrile (ACN).
11. Solution E: 0.1% formic acid in water.
12. Solvent F: 0.1% formic acid in ACN.
13. UniProt human proteomic database.
14. Masslynx Mass Spectrometry Software
® .
15. Progenesis
® QI for Proteomics version 4.0+ (Waters
Corporation).
16. Cytoscape software.
17. Ingenuity Pathways Analysis software.
2.3 Sample
Preparation
and Digestion
1. Human serum or plasma samples, or other similar samples.
2. 0.5- and 1.5-mL Eppendorf LoBind tubes.
3. 50 mM ammonium bicarbonate solution (pH 7.5).
4. Centrifugal Filter Units for 4-mL sample with 3000 Da molecular weight cutoff (MWCO) (Amicon
® Ultra-4 Centrifugal
Filter Units).
5. Refrigerated bench-top centrifuge with rotor for 15-mL conical tubes.
6. Spectrophotometer with a UV lamp (see Note 1).
7. 0.2% solution of RapiGestTM SF Surfactant (Waters
Corporation).
8. 100 mM dithiothreitol (DTT) in 50 mM ammonium bicarbonate solution (pH 7.5).
9. 300 mM iodoacetamide (IAA) in 50 mM ammonium bicarbonate solution (pH 7.5).
10. Sequencing-grade modified trypsin (Promega Corporation).
11. Trypsin resuspension dilution buffer (Promega Corporation).
12. Heater or thermoshaker.
13. 5% trifluoroaceticacid (TFA).
14. MassPREP™ Enolase digestion standards (ENO) (Waters
Corporation).
Human Blood Plasma Investigation Employing 2D UPLC-UDMS
E Data. . .
157
(100 A ˚ , 5 μm, 180 μm  20 mm) (Waters Corporation).
6. Acquity UPLC M-Class HSS T3 Column (1.8 μm,
75 μm  150 mm) (Waters Corporation).
7. NanoLock Spray dual electrospray ion source (Waters
Corporation).
8. 100 fmol/μL human [Glu
1 ]-fibrinopeptide B human
(Glu-Fib).
9. Solvent C: 20 mM ammonium formiate (pH 10).
10. Solvent D: 100% acetonitrile (ACN).
11. Solution E: 0.1% formic acid in water.
12. Solvent F: 0.1% formic acid in ACN.
13. UniProt human proteomic database.
14. Masslynx Mass Spectrometry Software
® .
15. Progenesis
® QI for Proteomics version 4.0+ (Waters
Corporation).
16. Cytoscape software.
17. Ingenuity Pathways Analysis software.
2.3 Sample
Preparation
and Digestion
1. Human serum or plasma samples, or other similar samples.
2. 0.5- and 1.5-mL Eppendorf LoBind tubes.
3. 50 mM ammonium bicarbonate solution (pH 7.5).
4. Centrifugal Filter Units for 4-mL sample with 3000 Da molecular weight cutoff (MWCO) (Amicon
® Ultra-4 Centrifugal
Filter Units).
5. Refrigerated bench-top centrifuge with rotor for 15-mL conical tubes.
6. Spectrophotometer with a UV lamp (see Note 1).
7. 0.2% solution of RapiGestTM SF Surfactant (Waters
Corporation).
8. 100 mM dithiothreitol (DTT) in 50 mM ammonium bicarbonate solution (pH 7.5).
9. 300 mM iodoacetamide (IAA) in 50 mM ammonium bicarbonate solution (pH 7.5).
10. Sequencing-grade modified trypsin (Promega Corporation).
11. Trypsin resuspension dilution buffer (Promega Corporation).
12. Heater or thermoshaker.
13. 5% trifluoroaceticacid (TFA).
14. MassPREP™ Enolase digestion standards (ENO) (Waters
Corporation).
Human Blood Plasma Investigation Employing 2D UPLC-UDMS
E Data. . .
157