Agilent 1100 Series nanoflow LC/MS system is a multidimensional liquid chromatography system for separating protein digests. This document describes the system configuration and usage methods
This document provides a technical overview of multidimensional liquid chromatography (MDLC) using the Agilent 1100 Series nanoflow LC/MS system. It details instrument configurations suitable for separating complex protein digests, covering both on-line and off-line two-dimensional nano LC/MS methods as well as one-dimensional enrichment techniques.
The text describes recommended equipment setups, including specific pumps, autosamplers, valves, and mass spectrometers like the LC/MSD Trap XCT. It outlines the principles of operation for different separation mechanisms, such as strong cation exchange (SCX) followed by reversed-phase chromatography, and lists compatible ZORBAX columns for these applications.
| Recommended Mass Spectrometer | LC/MSD Trap XCT (G2451AA) |
|---|---|
| Recommended Ion Source | Orthogonal nanospray source (G1982B) |
| Recommended SCX Column 1 | ZORBAX BioSCX Series IT, 35 x 0.38 mm |
| Recommended SCX Column 2 | ZORBAX BioSCX Series IT, 50 x 0.8 mm |
| Recommended C18 Column 1 | ZORBAX 300SB-C18, 150 mm x 75 µm, 3.5 µm particles |
| Recommended C18 Column 2 | ZORBAX 300SB-C18, 50 mm x 75 µm, 3.5 µm particles |
| Recommended C18 Trap Column | ZORBAX 300SB-C18, 5 x 0.38 mm, 5 µm particles |
The document recommends dissolving the 500-pmol bovine serum albumin (BSA) digest.
It is well-suited for extremely complex proteome samples such as whole cell lysates or tissues.