Proteomics & Protien Expression

Integrated Microcapillary HPLC

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Selected Protocol from "Proteins and Proteomics: A Laboratory Manual", David R. Goodlett, Eugene C. Yi, and Philippe Mottay.

Preparation and Use of an Integrated Microcapillary HPLC Column and ESI Device for Proteomic Analysis

David R. Goodlett, Eugene C. Yi, and Philippe Mottay

Implementation of separation techniques in miniaturized formats on-line with high-performance mass spectrometers and the development of miniaturized sprayers as electrospray ionization (ESI) ion sources have reduced the amount of peptide required for complete and routine sequence characterization from several picomoles to a few femtomoles and below. Arguably, much of this gain in sensitivity is due to the combination of concentration-dependent type ionization devices such as ESI and on-line capillary separation devices of very small internal diameter (I.D.). This can be primarily attributed to a reduced mass flow rate of solvents and other background constituents into the ESI source, which allows for greater sample ionization efficiency. For reasons of robustness, most microcapillary HPLC (µLC) work is done with 75- or 100-µm I.D. capillary columns that clog less frequently than 50-µm I.D. capillary columns. The following protocol describes the preparation of an integrated C18-packed capillary column-ESI microspray device. In this protocol, a polyimide capillary tip tapered to ~5 µm is used to hold the C18-derivatized particles in place.  

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Products Available for this Protocol
Protocol Material Description Product #  Product Name Add to Cart
Buffers, Solutions, and Reagents      
Acetonitrile 34967 Acetonitrile
Acetic acid A3102 Acetic acid, meets EP, BP, USP testing specifications
Derivatized silica resin View our entire list of Silica Gels
Ethanol (70%) E7148 Ethanol, for molecular biology, 190 proof
Methanol M1775 Methanol, Absolute - Acetone free

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