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Accurate label-free quantification ensured by minimal RSD values for retention times
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Stable, pulsation-free gradients from 100 nL/min
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Advanced Flow Control™ with two flow sensors prior to mixing
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Split-free gradient mixing – no flow splitting
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Fast separations with typical peak widths: 3 - 5 sec (FWHM) |
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With Bruker Daltonik HCTultra
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Pre-column: Analytical column: Sample: Solvents:
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1 cm, 100 μm ID, C18, 5 μm 150 mm, 75 μm ID, C18, 100 Å, 3 μm 100 fmol tryptic digest of BSA A: 0.4% acetic acid, water
B: 0.4% acetic acid, acetonitrile
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Gradient:
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300 nl/min, 5 - 35 %B in 10 minutes
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Courtesy of Bruker Daltonik, Germany
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Low retention time RSD values for complex mixtures |
With Thermo Fisher LTQ Orbitrap XL FT scan 390-1500 m/z
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Column: Sample:
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15 cm, 75 µm ID, C18 5 µm 100 fmol annexin V
800 fmol gelsolin
300 fmol BSA digestion mixture
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Gradient:
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300 nl/min, 3 - 35 %B in 25 minutes
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Courtesy of J.M. Asara, Beth Israel Deaconess Medical Center, Boston, MA.
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Efficient Peltier-controlled sample cooling
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Injection volume selectable in 0.01 μl steps
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Injection needle washed outside and inside
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Custom wash with 2 extra solvent bottles
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Excellent sensitivity and reproducibility |
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With Thermo Fisher TSQ- Quantum Ultra
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MRM scan (m/z 651.3) 200 attomole BSA
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Injected sample: 2µl of 100 amol/µl BSA tryptic digest.
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RT RSD 0.08%
Courtesy of Dr.P. Taylor, Hospital for Sick Children, Ontario.
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Automated 2D (SCX-RPC) separation with a 1-column system configuration |
Base peak chromatograms
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With Thermo Fisher LTQ Orbitrap XL
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Autosampler within EASY-nLC used for precise step elution in SCX prior to RPC gradient elution and MS-MS analysis.
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Separation: One-column set up of EASY-nLC with a biphasic analytical column (5 cm of Phenomenex Luna SCX 5 µm + 5 cm of Michrom Magic C18 5 µm). After injection of treated mouse placental tissue, 2D separation was performed by injecting a salt gradient in 10 steps, from 0.05 M to 0.5 M ammonium acetate. Control 1D experiments were used to verify the performance.
Courtesy of Dr. T. Kislinger, Ontario Cancer Institute (OCI), Canada Extracted ion currents for 474.79 Th ion
2D experiments identified 3-4 times as many proteins as 1D control
| Scoring Criteria | 1D | 2D-1 | 2D-2 | Combined 2D |
Protein Prophet 95%
Peptide Prophet 95%
2 unique peptide minimum | 413 | 1220 | 1390 | 1604 |
Protein Prophet 95%
Peptide Prophet 95%
1 peptide minimum | 524 | 1461 | 1670 | 1906 |
X! Tandem e score < -2.0
2 unique peptide minimum | 476 | 1338 | 1540 | 1788 |
X! Tandem e score < -1.5
2 unique peptide minimum | 518 | 1439 | 1656 | 1907 |
X! Tandem e score < -2.0
1 peptide minimum | 788 | 2171 | 2354 | 2673 |
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