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Life Science > Proteomics > Recombinant Protein Expression > Protocols > Membrane-bound Peptide Arrays
Proteomics & Protien Expression

Membrane-bound Peptide Arrays

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

Synthesis and Probing of Membrane-bound Peptide Arrays

Ronald Frank
Department of Chemical Biology, GBF (German Research Center for Biotechnology)
38124 Braunschweig, Germany

Stefan Dubel
Institute for Biochemistry and Biotechnology, Technical University of Braunschweig
D-38106 Braunschweig, Germany

  Protein Protein Interactions

Buy this book, Z719145

 
ABSTRACT

The following protocol describes the synthesis of short linear peptides, or peptide pools, on modified cellulose membranes, and the detection of their protein-binding partners. Peptides are synthesized from their carboxyl termini using Fmoc-amino acid derivatives. After completion of the synthesis and cleavage of all side-chain protecting groups, the peptide array on the membrane is incubated with the potential interaction partners to identify their target sequences.


Products Available for this Protocol
Protocol Material Description Product #  Product Name Add to Cart
Buffers, Solutions, and Reagents      
Bromophenol blue indicator (BPB) B0126 Bromophenol Blue
   
Acetic acid A9967 Acetic acid
   
DMF D4551 N,N-Dimethylformamide
   
1-Methyl-2-pyrrolidinone (NMP) 494496 1-Methyl-2-pyrrolidinone
   
DIC (N,N'-Diisopropylcarbodiimide) D125407 N,N'-Diisopropylcarbodiimide
   
Acetic anhydride 539996 Acetic anhydride
   
Piperidine 571261 Piperidine
   
Methanol or ethanol, technical grade E7148 Ethanol
   
Trifluoroacetic acid (TFA, synthesis grade) T6508 Trifluoroacetic acid
   
Triisobutylsilane (TIBS) 278785 Triisobutylsilane
   
Water W4502 Water
   
Dichloro­ methane (DCM) 66738 Dichloromethane
   
Tris-buffered saline (TBS) T6664 Tris Buffered Saline
   
NaCl S3014 Sodium chloride
   
KCl P9541 Potassium chloride
   
Tris-base T6066 Trizma® base
   
HCl 26612 Hydrogen chloride
   
NaOH 72068 Sodium hydroxide solution
   
Tween-20 P9416 TWEEN® 20
   
Phosphate-buffered saline (PBS) P5368 Phosphate buffered saline
   
Na2HPO4•2H2O S9763 Sodium phosphate dibasic
   
KH2PO4 60353 Potassium phosphate dibasic
   
Citric acid C2404 Citric acid
   
Casein-based blocking buffer concentrate C7594 Casein blocking buffer
   
Sucrose S0389 Sucrose
   
Methanol M3641 Methanol
   
Sodium dodecyl sulfate (SDS) L3771 Sodium dodecyl sulfate
   
Alkaline phosphatase (AP)-conjugated detection antibodies Antibody Explorer Antibodies  
Tris-HCl (pH 7.6) T5941 Trizma® hydrochloride
   
Glycine 50046 Glycine
   
Horseradish peroxidase (HRP)-conjugated detection antibodies Antibody Explorer Antibodies  
Urea U5378 Urea
   
Mercaptoethanol M3148 2-Mercaptoethanol
   

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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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