From sequence to antibody: genetic immunisation is suitable to generate antibodies against a rare plant membrane protein, the KAT 1 channel.
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.
Gehwolf
Renate
R
Molecular Plant Physiology, Division of Allergy and Immunology, Department of Molecular Biology, University of Salzburg, Salzburg, Austria.
Weiss
Richard
R
Gabler
Maximilian
M
Hurst
Annette C
AC
Bertl
Adam
A
Thalhamer
Josef
J
Obermeyer
Gerhard
G
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
D000911:Antibodies, Monoclonal; D029681:Arabidopsis Proteins; C501210:KAT1 protein, Arabidopsis; D024661:Potassium Channels, Inwardly Rectifying; D011994:Recombinant Proteins; D019444:Vaccines, DNA