Download Combinatorial Peptide Library Protocols (Methods in by Shmuel Cabilly PDF

By Shmuel Cabilly

A important selection of new and specific strategies for the development and use of peptide libraries. those robust equipment contain protocols for the chemical synthesis of peptide libraries, for developing peptide libraries which are displayed at the floor of filamentous phage or micro organism, and for the quick screening of those libraries for molecules with biospecific houses. extra equipment enable selecting particular enzyme substrates, investigating the popularity spectra of varied binding proteins, epitope mapping, and picking mimotopes. state of the art innovations, mixed with transparent step by step directions, make this e-book a vital instrument within the number of peptides appropriate for drug improvement.

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Additional info for Combinatorial Peptide Library Protocols (Methods in Molecular Biology Vol 87)

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1). On the first column synthesize the onginal peptide, on the other columns substitute each position of the peptide by all 20 amino acids m alphabetical order. Carry out the peptide synthesis steps as described in the previous sections. Here are some recommendations for the pipeting order. 9. Synthesis of a Pepticfe Scanning Library The first step is the derivation of the peptrde sequences from the protein sequence. A peptide length of 12 amino acids and an overlap of 9 amino acids is recommended In Fig.

11. ) have to be optimized for each system. In general, if you do not get any srgnal or your signals are too weak there are several posstbilities to optimize: a. Increase the ligand and/or antibody concentrations. b Prolong the incubation time with ligand to overnight at 4°C. c. Prolong the mcubation time with secondary antibody to 3 h. d. Shorten the washing times, use washing buffer without Tween-20. e. Shorten the blockmg time to 3 h. If the background IS too high, try the followmg changes: a.

38 Kramer and Schneider-Mergener Acknowledgments This work was supported by the BMBF, DFG, and Fonds der Chemlschen Industrie. We thank Beret Hoffmann and Christlane Landgraf for excellent technical assistance. References 1 Frank, R (1992) Spot synthesis. an easy technique for the positionally addressable, parallel chemical synthesis on a membrane support Tetrahedron 48, 9217-9232. 2. , Volkmer-Engert, R , Maim, R , Remeke, U , and SchneiderMergener, J (1993) Simultaneous synthesis of peptide libraries on single resin and contmuous membrane supports: identification of protem, metal and DNA bmdmg peptide mixtures.

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