The Xenopus oocyte as an ectopic expression system for the selection of protein isoform-specific antibodies.
A panel of Xenopus oocytes, each injected with cRNA coding for one specific isoform of the rat brain RCK family of voltage gated potassium channel proteins, was employed to screen for isoform-specific monoclonal antibodies. Several days after injection, cryosections of embedded oocytes were produced and were employed in immunohistochemical analysis of antibody binding. Of the advantageous properties of the assay, it employs the native antigen, it can be applied to homooligomeric and heterooligomeric proteins, and cryosections of the same batch can be stored frozen for later tests. The method may be advantageous also for the selection of isoform-specific antibodies of other protein families.
Reinhardt-Maelicke
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Institut für Physiologische Chemie, Johannes Gutenberg-Universität, Mainz, Germany.
Kurz
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Sewing
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Stocker
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Pongs
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O
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D005260:Female; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009419:Nerve Tissue Proteins / Q000276:immunology; D009865:Oocytes / Q000378:metabolism*; D015221:Potassium Channels / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology; D014982:Xenopus laevis
D000911:Antibodies, Monoclonal; D009419:Nerve Tissue Proteins; D015221:Potassium Channels; D011993:Recombinant Fusion Proteins