 On-line 2D Nano LC/MS is well suited for digests from gel spots or bands which need further separation and for subcellular proteome fractions of higher complexity. It is also useful for identifying biomarkers in pre-enriched biological fluids like blood, spinal fluid or urine And it is the best choice for further separation of affinity-purified post-translationally modified (PTM) peptides or protein complexes for functional proteomics research. Here, Agilent offers a flexible approach which is based on the Agilent Nanoflow Proteomics Solution.
 Recommended configuration for a conventional 2D Nano LC/MS system
- Agilent 1260 Infinity Nanoflow LC system for MS (G2229A) - Includes nanoflow pump, micro vacuum degasser, micro well-plate autosampler, and handheld controller. Optimum solution to work at stable, precise nanoliter flow rates in combination with nanoelectrospray for the highest sensitivity in your proteomics work
- Agilent 1200 Series 2-position/10-port micro valve (G1163A) - offers highest flexibility for different techniques in the proteomics area
- Agilent 1260 Infinity Capillary Pump - for robust and reliable flow in the µL range
- Agilent 6500 Series Accurate Mass Q-TOF LC/MS - offers highest sensitivity with an extended capacity trap
- Orthogonal nanoelectrospray source (G1982B) - for robust and easy-to-use nanospray
- Nanoflow LC start-up kit (G2228-68700) - includes nano RP and RP enrichment columns and optimized fittings and capillaries that make handling easy
- Agilent 1200 Infinity Series Column Organizer (G1383A) - to adjust valves correctly to minimize the capillary length between the LC system and MS system
- Spectrum Mill MS Proteomics Workbench (G2721A)- for proteomics data analysis
- Thermostat (G1330B) for Agilent 1260 Infinity Autosampler, Well-plate Sampler and Fraction Collector - to prevent degradation of thermolabile samples
Recommended chromatographic columns
- Ion Exchange column(s) for the 1st dimension of 2D LC
ZORBAX BioSCX Series II, 300 µm × 35 mm, 3.5 µm; strong cation exchanger (5065-9912)
ZORBAX BioSCX Series II, 0.8 mm × 50 mm, 3.5 µm; strong cation exchanger (5065-9942)
- Reversed-phase column for peptide enrichment before 2nd dimension of 2D LC:
ZORBAX 300SB-C18, 300 µm × 5 mm, 5 µm (5/pk) (included in G2228A Nanoflow LC start-up kit) (5065-9913)
- Reversed-phase columns for 2nd dimension of 2D LC:
ZORBAX 300SB-C18, 75 µm × 150 mm, 3.5 µm (included in G2228A Nanoflow LC start-up kit) (5065-9911)
- Holder for 5-mm trap cartridges (5065-9915)
Recommended configuration for an Agilent 1260 Infinity HPLC-Chip/MS on-line 2D nano LC/MS system
- Agilent 1260 Infinity Nanoflow LC system for MS (G2229A) - Includes nanoflow pump, micro vacuum degasser, micro well-plate autosampler, and handheld controller. Optimum solution to work at stable, precise nanoliter flow rates in combination with nanoelectrospray for the highest sensitivity in your proteomics work
- Agilent 1260 Infinity HPLC-Chip/MS system (G4240A) - fully integrated HPLC-Chip offers easy to use, robust nanospray LC-MS for proteomics applications.
- Agilent 1260 Infinity Capillary Pump with micro vacuum degasser (G1382A) - for robust and reliable flow in the µL range
- Agilent 6500 Series Accurate Mass Q-TOF LC/MS - offers highest sensitivity with an extended capacity trap
- Spectrum Mill MS Proteomics Workbench (G2721A)- for proteomics data analysi
- Thermostat for Agilent 1260 Infinity Autosampler, Well-plate Sampler and Fraction Collector (G1330B) - to prevent degradation of thermolabile samples
Recommended HPLC-Chips and 1st dimension LC columns
- Ion Exchange column(s) for the 1st dimension of 2D LC
ZORBAX BioSCX Series II, 300 µm × 35 mm, 3.5 µm; strong cation exchanger (5065-9912)
ZORBAX BioSCX Series II, 0.8 mm × 50 mm, 3.5 µm; strong cation exchanger (5065-9942)
- Protein ID #1 HPLC-Chip - for low-medium complexity samples. Integrated enrichment column, separation column, capillaries and spray emitter directly on the chip. 40nl enrichment column and 43 mm x 75um analytical column packed with ZORBAX 300SB C-18 5um. (G4240-62001). ZORBAX 300SB-C18, 300 µm × 5 mm, 5 µm (5/pk) (included in G2228A Nanoflow LC start-up kit) (5065-9913)
- Protein ID #2 HPLC-Chip - for medium-high complexity samples. Integrated enrichment column, separation column, capillaries and spray emitter directly on the chip. 40-nL enrichment column and 150 mm x 75µm analytical column packed with ZORBAX 300SB C-18 5 µm (G4240-62002)
Back to Multidimensional LC Separation of Protein Digests
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