Anti-c-myc antibody 9E10: epitope key positions and variability characterized using peptide spot synthesis on cellulose. |
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| Authors: | K Hilpert G Hansen H Wessner G Küttner K Welfle M Seifert W H?hne |
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| Institution: | Institut für Biochemie, Universit?tsklinikum Charité, Humboldt-Universit?t zu Berlin, Monbijoustr. 2, 10117 Berlin, Germany. |
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| Abstract: | The 9E10 antibody epitope (EQKLISEEDL) derives from a protein sequence in the human proto-oncogen p62(c-myc) and is widely used as a protein fusion tag. This myc-tag is a powerful tool in protein localization, immunochemistry, ELISA or protein purification. Here, we characterize the myc-tag epitope by substitutional analysis and length variation using peptide spot synthesis on cellulose. The key amino acids of this interaction are the core residues LISE. The shortest peptide with a strong binding signal is KLISEEDL. Dissociation constants of selected peptide variants to the antibody 9E10 were determined. scFv constructs with the shortest possible myc-tags were successfully detected by Western blot and ELISA, giving a signal comparable to that of the original myc-tag. |
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