|
|
The following Cold Spring Harbor Laboratory Press protocols are brought to you by BioSupplyNet in a partnership with Sigma-Aldrich. A list of products is available to use in each protocol.
View the protocol on BioSupplyNet.com. |
|
Selected Protocol from "Proteins and Proteomics: A Laboratory Manual", Richard J. Simpson.
Preparation and Use of an Integrated Microcapillary HPLC Column and ESI Device for Proteomic Analysis
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.
|
Buy this book, Z700428
|
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 |
|
back to top
View all available Cold Spring Harbor Laboratory Press
Protocols
Product Association Disclaimer:
The Sigma-Aldrich products listed for this specific protocol were selected either to match or to
supplement the products listed within the actual protocol. The products/reagents from Sigma-Aldrich
have been qualified for usage, but may not have been validated for this specific application. Please
refer to the detailed product description on the usage of specific products of interest.
|
