title: From sequence to antibody: genetic immunisation is suitable to generate antibodies against a rare plant membrane protein, the KAT 1 channel.
abstract: Monoclonal antibodies against the K(+) channel KAT1 of Arabidopsis thaliana, a low abundance, plant plasma membrane protein, were generated by genetic immunisation to avoid the time and labour consuming purification of native or recombinant proteins and peptides usually necessary for conventional immunisation techniques. The resulting polyclonal and monoclonal antibody sera recognised a single protein band in a microsomal fraction of wild-type A. thaliana leaves and in membrane fractions of transgenic yeast cells and tobacco plants expressing the KAT1 protein. Therefore, genetic immunisation is suitable for generating monoclonal antibodies against plant proteins and particularly, against plant membrane proteins of low abundance.
authors.affiliation: Molecular Plant Physiology, Division of Allergy and Immunology, Department of Molecular Biology, University of Salzburg, Salzburg, Austria.
authors.forename: Renate
authors.initials: R
authors.lastname: Gehwolf
authors.forename: Richard
authors.initials: R
authors.lastname: Weiss
authors.forename: Maximilian
authors.initials: M
authors.lastname: Gabler
authors.forename: Annette C
authors.initials: AC
authors.lastname: Hurst
authors.forename: Adam
authors.initials: A
authors.lastname: Bertl
authors.forename: Josef
authors.initials: J
authors.lastname: Thalhamer
authors.forename: Gerhard
authors.initials: G
authors.lastname: Obermeyer
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D017360:Arabidopsis / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000276:immunology*; D005260:Female; D005822:Genetic Vectors; D006825:Hybridomas / Q000276:immunology; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000235:genetics; D024661:Potassium Channels, Inwardly Rectifying / Q000235:genetics* / Q000276:immunology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014026:Nicotiana / Q000235:genetics; D019444:Vaccines, DNA / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D029681:Arabidopsis Proteins; C501210:KAT1 protein, Arabidopsis; D024661:Potassium Channels, Inwardly Rectifying; D011994:Recombinant Proteins; D019444:Vaccines, DNA