Intradermal DNA Immunization: Antisera Specific for the Membrane Lectin MR60/ERGIC-53 |
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Authors: | Violaine Carrière Ludovic Landemarre Valérie Altemayer Geneviève Motta Michel Monsigny Annie-Claude Roche |
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Affiliation: | (1) Glycobiologie, Centre de Biophysique Moléculaire, CNRS, Rue Charles Sadron, F-45071 Orléans cedex 02, France;(2) Agro-Bio, La Chavannerie, rue Denis Papin, F-45240 La Ferté, Saint Aubin;(3) Laboratoire de Parasitologie, UFR des Sciences Université d'Orléans, BP 6759, F-45067 Orléans cedex 2 |
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Abstract: | The trafficking of intracellular membrane proteins in Golgi apparatus, endoplasmic reticulum or intermediate compartment has not yet been fully elucidated. The human MR60/ERGIC-53 and the rat p58 proteins are one such protein; and to study them in cell-free and in situ systems, high quality monospecific antisera are required. Highly specific antisera have been obtained after immunization of mice with plasmids containing a gene encoding either the full length or a truncated protein. The best results were obtained after intradermal injections of a plasmid encoding a truncated protein comprising both the luminal carbohydrate recognition domain and the stem down to a cysteine residue close to the C-terminal end, but neither the transmembrane nor the cytosolic domains. Such antisera have a very high titer and are very efficient tools to visualize the MR60 protein in situ or to selectively precipitate the MR60 proteins from a whole cell lysate. |
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Keywords: | Confocal microscopy ERGIC-53 Human lectin Mannose-specific lectin Mouse antibody MR60 |
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