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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
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. |
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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 |
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| 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 |
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| 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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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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