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Life Science > Proteomics > Recombinant Protein Expression > Protocols > Integrated Microcapillary HPLC
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Proteomics & Protien Expression
Integrated Microcapillary HPLC | ||
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Sigma® has prepared product lists for commonly used protocols from Cold Spring Harbor Protocols. |
View the entire protocol on BioSupplyNet.com.
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Selected Protocol from "Proteins and Proteomics: A Laboratory Manual", David R. Goodlett, Eugene C. Yi, and Philippe Mottay. David R. Goodlett, Eugene C. Yi, and Philippe Mottay |
| ABSTRACT | ||
| 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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