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HPLC separation and electrospray ionization have proven an effective combination for analyzing complex mixtures of biomolecules. The sensitivity of both techniques is improved primarily by scaling down their size. We employ an Agilent nanopump flowing at 200-300 nanoliters/minute, a 75 um ID C18 column, and an emitter with an aperture of 8 um to obtain high sensitivity peptide identifications. With the addition of a capillary pump and a switching valve, we have the option of performing both one and two-dimensional chromatography experiments for the analysis of extremely complex mixtures such as cell lysates, organelle contents, or subcellular fractions. Our nanospray source is interfaced to the Agilent XCT Plus ion trap.
For excellent information on nanospray, please visit the New Objective website at www.newobjective.com.
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