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Proteomics & Protein Expression
Screening Kinase Phosphorylation Protocol
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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.
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| View the entire protocol on BioSupplyNet.com. |
| Selected Protocol from "Protein-Protein Interactions", 2nd Edition, E. Golemis, P. Adam. |
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Screening Kinase Phosphorylation Motifs Using Peptide Libraries
Isaac A. Manke and Michael B. Yaffe
Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, MA
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Buy this book, Z719145
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| ABSTRACT |
| Determining which protein kinases are responsible for phosphorylating which protein substrates is critical for mapping signal transduction cascades that regulate cell growth, division, differentiation, and death. Oriented peptide library screening, combined with bioinformatics-based searches of protein sequence databases, offers a strategy for identifying substrates of particular protein kinases. |
| Products Available for this Protocol |
| Protocol Material Description |
Product # |
Product Name |
Add to Cart |
| Acetic acid, 0.1 m in water |
45754 |
Acetic acid solution |
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| Acetic acid, 30% in water |
45754 |
Acetic acid solution |
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| Acetone |
00570 |
Acetone |
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| Acetonitrile |
92670 |
2,4,4-Trimethyl-1-pentene |
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| Ammonium acetate, 0.1% (pH 11.5), freshly prepared and stored at 4°C |
A1542 |
Ammonium acetate |
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| Ammonium acetate, 0.1% (pH 8.0), freshly prepared and stored at 4°C |
A1542 |
Ammonium acetate |
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| Ammonium acetate, 0.1% (pH 9.5), freshly prepared and stored at 4°C |
A1542 |
Ammonium acetate |
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| ATP, 10 mm (pH 7.0) (store at –20°C) |
A6559 |
Adenosine 5'-triphosphate disodium salt solution |
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| DEAE Sephacel (Sigma I6505) |
I6505 |
Diethylaminoethyl–Sephacel™ |
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| Degenerate Ser- or Thr-containing peptide libraries |
Custom Peptides |
Custom Peptides |
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| EDTA, 0.1 m (pH 8.0) |
E7889 |
Ethylenediaminetetraacetic acid disodium salt solution |
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| FeCl3, 100 mm in 0.1 m acetic acid (prepare fresh) |
451649 |
Iron(III) chloride |
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| Immobilized iminodiacetic acid agarose (IDA agarose) with 1,4 butanediol diglycidyl ether spacer |
I4758 |
Iminodiacetic acid Agarose |
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| Methanol, dry |
M3641 |
Methanol |
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| Phosphoric acid, 85% |
452289 |
Phosphoric acid |
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| Solution 1: 50 mm 2-(N-morpholino)ethanesulfonic acid (MES) (pH 5.5) containing 1 m NaCl |
M3671 |
MES |
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| Standard reagents for peptide synthesis and protein sequencing (if this is being done in-house) |
Standard Reagents |
Standard reagents for peptide synthesis and protein sequencing |
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| Thionyl chloride |
447285 |
Thionyl chloride |
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| 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. |
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