PRODUCTION AND CHARACTERIZATION OF AN ANTIBODY-DIRECTED AGAINST THE MOUSE 5HT1B RECEPTOR

We have developed a polyclonal antibody directed against a peptide loc ated in the third intracellular loop of the 5HT1B receptor. Its charac terization was carried out using NIH cells stably transfected with a e ukaryotic expression vector containing the mouse 5HT1B receptor cDNA. The synthetic antigenic peptide had a unique sequence to the mouse 5HT 1B receptor corresponding to amino acids 273-287 localized in the thir d intracellular loop. In dot blot analysis, antisera detected 2 ng to 2 mu g of synthetic peptide at dilutions of 1/200-1/20 000 and bound a ntibody was visualized using an immunoperoxidase procedure. Preimmune serum showed no immunoreactivity to the synthetic peptide. NIH cells s tably transfected expressing mouse 5HT1B receptor displayed an intense immunoreactivity with the antiserum. In contrast no immunoreactivity was seen in any of the control experiments. In the present study, we h ave produced a specific antibody which is an essential tool suitable f or immunocytochemical applications such as regional distribution, anat omical localization and phenotypical characterization of the cells exp ressing the 5HT1B receptors in brain by double immunolabelling procedu re.