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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.
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Selected Protocol from "Proteins and Proteomics: A Laboratory Manual", Richard J. Simpson.
Analysis of Complex Protein Mixtures Using Multidimensional Protein Identification Technology (MuDPIT)
Richard J Simpson
ABSTRACT
This protocol describes the analysis of a proteome using multidimensional protein identification technology (MuDPIT), which couples 2D-LC to MS/MS, to resolve and identify peptides from complex mixtures. In this method, a pulled capillary microcolumn is packed with two independent chromatography phases: a strong cation exchanger and reversed-phase matrix material. Once a complex peptide mixture is loaded onto the system, no additional sample handling is required, because, as the peptides elute from the column, they are directed into the ESI ion-trap mass spectrometer, where they are ionized, mass-selected, and fragmented. Finally, advanced search algorithms match the fragmented peptides to their respective proteins in a database. |
Buy this book, Z700428
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Products Available for this Protocol
| Protocol Material Description |
Product # |
Product Name |
Add to Cart |
| Buffers, Solutions, and Reagents |
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| Acetonitrile |
34967 |
Acetonitrile |
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| Ammonium bicarbonate |
A6141 |
Ammonium bicarbonate |
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| Trypsin |
T6567 |
Trypsin from porcine pancreas, Proteomics Grade,
Dimethylated |
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| Methanol |
M1775 |
Methanol, Absolute - Acetone free |
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| Dithiothreitol (DTT) |
D9779 |
DL-Dithiothreitol, for molecular biology |
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| Urea |
U5378 |
Urea, for molecular biology |
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| Heptafluorobutyric acid |
77247 |
Heptafluorobutyric acid |
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| Calcium chloride |
21115 |
Calcium chloride solution |
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| Endoproteinase Lys-C |
P3428 |
Endoproteinase Lys-C from Lysobacter enzymogenes |
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| Iodoacetamide (IAA) |
I1149 |
Iodoacetamide |
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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.
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