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Life Science > Proteomics > Recombinant Protein Expression > Protocols > Screening Kinase Phosphorylation Protocol
Proteomics & Protein Expression

Screening Kinase Phosphorylation Protocol

cshlp-small Sigma® has prepared product lists for commonly used protocols from Cold Spring Harbor Protocols.

View the entire protocol on BioSupplyNet.com.

Selected Protocol from "Protein-Protein Interactions", 2nd Edition, E. Golemis, P. Adam.

Screening Kinase Phosphorylation Motifs Using Peptide Libraries

Isaac A. Manke and Michael B. Yaffe
Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, MA

  Protein Protein Interactions

Buy this book, Z719145

 
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
   
Acetic acid, 30% in water 45754 Acetic acid solution
   
Acetone 00570 Acetone
   
Acetonitrile 92670 2,4,4-Trimethyl-1-pentene
   
Ammonium acetate, 0.1% (pH 11.5), freshly prepared and stored at 4°C A1542 Ammonium acetate
   
Ammonium acetate, 0.1% (pH 8.0), freshly prepared and stored at 4°C A1542 Ammonium acetate
   
Ammonium acetate, 0.1% (pH 9.5), freshly prepared and stored at 4°C A1542 Ammonium acetate
   
ATP, 10 mm (pH 7.0) (store at –20°C) A6559 Adenosine 5'-triphosphate disodium salt solution
   
DEAE Sephacel (Sigma I6505) I6505 Diethylaminoethyl–Sephacel™
   
Degenerate Ser- or Thr-containing peptide libraries Custom Peptides Custom Peptides  
EDTA, 0.1 m (pH 8.0) E7889 Ethylenediaminetetraacetic acid disodium salt solution
   
FeCl3, 100 mm in 0.1 m acetic acid (prepare fresh) 451649 Iron(III) chloride
   
Immobilized iminodiacetic acid agarose (IDA agarose) with 1,4 butanediol diglycidyl ether spacer I4758 Iminodiacetic acid Agarose
   
Methanol, dry M3641 Methanol
   
Phosphoric acid, 85% 452289 Phosphoric acid
   
Solution 1: 50 mm 2-(N-morpholino)ethanesulfonic acid (MES) (pH 5.5) containing 1 m NaCl M3671 MES
   
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  
Thionyl chloride 447285 Thionyl chloride
   

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