By Ronald Frank, Jens Schneider-Mergener (auth.), Joachim Koch, Michael Mahler (eds.)
Proteins interacting with diversified ligands - proteins, peptides or DNA - are the fundamental rules underlying many organic procedures, similar to antigen-antibody binding, sign transduction or receptor binding.
The means of oligopeptide synthesis on a cellulose membrane and the next binding assays enable the research of protein interactions. a selected benefit of those peptide arrays (SPOT - know-how) is the excessive variety of oligopeptide probes that may be demonstrated in parallel. specific protocols for peptide synthesis, and the research of protein-protein, protein-DNA interactions in addition to epitope mapping are awarded during this guide. it really is ultimate not just for uncomplicated examine laboratories but in addition for diagnostic and healing functions on account that many illnesses are concerning dysfunctions in protein popularity and binding.
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Extra info for Peptide Arrays on Membrane Supports: Synthesis and Applications
By following the stripping conditions (SDS, urea, acetic acid) described, these membranes can be reused 50 times and more (Frank and Owervin 1996). However, also with these membranes it is not advisable to use strongly acidic stripping conditions. The limited solvent compatibility of the cellulose membranes needs to be considered only if the peptides are to be modified during or after synthesis in solvents different from DMF, 1-methyl-2-pyrrolidone or water. Polar solvents are necessary to ensure a complete reaction while other common, less polar organic solvents like pyridine, THF or dichloromethane often lead to a low conversion.
In: Epton R (ed) Proc Int Symp on Innovation and Perspectives in Solid Phase Synthesis, Oxford, August 1993. Mayflower Worldwide Ltd, Birmingham,pp 509-512 Frank R, Giiler S (1990) Verfahren zur schnellen Synthese von tragergebundenen oder freien Peptiden oder Oligonucleotiden, damit hergestelltes 21 22 RoNALD FRANK, JENS ScHNEIDER-MERGENER Flachmaterial, dessen Verwendung sowie Vorrichtung zur Durchfiihrung des Verfahrens. 9 Frank R, Giiler S, Krause S, Lindenmaier W (1991) Facile and rapid spotsynthesis of large numbers of peptides on membrane sheets.
9. Wash off excess acetic anhydride with DMF 10. Start next cycle with step 2, or, after the last cycle, proceed with step 11. End ofthe synthesis 11. Perform a final Fmoc deprotection (steps 2 and 3). 12. Wash off excess piperidine with DMF and stain with BPB (step 4). 13. Acetylate the terminal amino groups with a mixture of 2 o/o (v/v) acetic anhydride in DMF (step 8). 14. Wash off excess acetic anhydride with DMF. 15. Wash membrane with ethanol and dry. 16. Cleave two times for 1 h with TFA/dichloromethane/water/ diisobutylsilane 50:50:2:3 (v/v).