|
|
Proteomics & Protien Expression
Labeling of Quantitation
|
|
|
 |
The following Cold Spring Harbor Laboratory Press protocols are brought to you by BioSupplyNet in an exclusive partnership with Sigma-Aldrich. Each month, 2 new protocols will be featured, each with a list of products available from Sigma-Aldrich.
|
|
View the entire protocol on BioSupplyNet.com.
|
ABSTRACT
Stable isotope coding strategies are of immense value in determining protein concentration changes in cells and tissues triggered by regulatory stimuli (e.g., drugs and toxins) and disease (e.g., caused by mutational changes). Recognizing and identifying the small number of proteins whose expression levels differ as a consequence of disease or external stimuli are complicated by the complexity of biological extracts and the fact that protein posttranslational modifications are numerous and can occur at many sites on a protein.
One way of quantifying global protein expression patterns involves in vivo labeling. Using the following protocol, stable isotopes can be incorporated into metabolic products, and the relative difference between these products from cells grown in normal or isotope-enriched media can be readily quantified by MS analysis. This protocol was provided by Yoshiya Oda (Eisai Co., Ltd., Ibaraki, Japan). |
|
Buy this book, Z700428
|
| Products Available for this Protocol |
| Protocol Material Description |
Product # |
Product Name |
Add to Cart |
| Buffers, Solutions, and Reagents |
|
|
|
| Acetone |
323772 |
Acetone |
|
| Acetonitrile |
34967 |
Acetonitrile |
|
| Ammonium bicarbonate |
A6141 |
Ammonium bicarbonate |
|
| Trypsin |
T6567 |
Trypsin from porcine pancreas, Proteomics Grade, Dimethylated |
|
| H2O |
W4502 |
Water, Molecular Biology Reagent |
|
| 2-Mercaptoethanol |
M6250 |
2-Mercaptoethanol, >99% |
|
| Methanol |
M1775 |
Methanol, Absolute - Acetone free |
|
| Acetic Acid |
A6283 |
Acetic acid, >99% |
|
| Potassium ferricyanide |
P8131 |
Potassium hexacyanoferrate(III), 99% |
|
| Protein extraction reagent |
Protein Extraction Reagents |
Protein Extraction Reagents |
|
| Protease inhibitor cocktail |
Protease inhibitor cocktail |
Protein Extraction Reagents |
|
| Sodium thiosulfate |
S7026 |
Sodium thiosulfate |
|
| Tris-Cl |
T5941 |
Trizma® hydrochloride, for molecular biology, Biotechnology Performance Certified, cell culture tested |
|
| Urea |
U5378 |
Urea, for molecular biology |
|
| Triton X-100 |
T8787 |
Triton® X-100, for molecular biology |
|
| Dithiothreitol (DTT) |
D9779 |
DL-Dithiothreitol, for molecular biology |
|
| Ampholine |
Carrier Ampholytes |
Carrier Ampholytes |
|
| Trichloroacetic acid (TCA) |
T8657 |
Trichloroacetic acid, for electrophoresis |
|
| SDS-PAGE sample buffer (2x sample buffer for SDS-PAGE) |
S9788 |
2× SDS Sample Buffer |
|
| Glycerol |
G5516 |
Glycerol, for molecular biology |
|
| SDS |
L4390 |
Sodium dodecyl sulfate, for molecular biology |
|
| Bromophenol blue |
B0126 |
Bromophenol blue |
|
| Triglycine |
G1377 |
Gly-Gly-Gly |
|
| TFA |
299537 |
Trifluoroacetic acid, >99%, purified by redistillation, for protein sequencing |
|
| Cell culture growth media |
Classic Media and Salts |
Classic Media and Salts |
|
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. |
| |
|
