RESUMO
Peptides corresponding to sequences derived from predicted extra- and intracellular loops of the rat bradykinin receptor were analyzed for interspecies homology as well as for matches within the present dataset of protein sequences to provide a theoretical basis for the specific recognition of the native cognate protein by antibodies raised against these antigens. Application of polyclonal antibodies raised against the selected peptides allowed the immunocytochemical localization of the native receptor protein in cells of rat and human origin. The detection of the molecule was achieved by different immunohisto- and immunocytochemical methods in combination with light, fluorescence, confocal optical laser and electron microscopy. These results were compared to localization studies by autoradiography. Distribution and subcellular localization were determined in human neutrophils, human epithelial carcinoma cells (A431) and in rat kidney tissue.
Assuntos
Cininas/fisiologia , Receptores da Bradicinina/metabolismo , Sequência de Aminoácidos , Animais , Autorradiografia , Sítios de Ligação de Anticorpos , Células Cultivadas , Técnicas de Cultura , Imunofluorescência , Humanos , Técnicas Imunoenzimáticas , Rim/metabolismo , Dados de Sequência Molecular , Neutrófilos/metabolismo , Ratos , Ratos Sprague-Dawley , Receptor B2 da Bradicinina , Homologia de Sequência , Especificidade da Espécie , Células Tumorais CultivadasRESUMO
Peptides corresponding to sequences derived from predicted extra- and intracellular loops of the rat bradykinin receptor were analyzed for interspecies homology as well as for matches within the present dataset of protein sequences to provide a theoretical basis for the specific recognition of the native cognate protein by antibodies raised against these antigens. Apllication of polyclonal antibodies raised against the selected peptides allowed the immunocytochemical localization of the native receptor protein in cells of rat and human origin. The detection of the molecule was achieved by different immunohisto- and immunocytochemical methods in combination with light, fluorescence, confocal optical laser and electron microscopy. These results were compared to localization studies by autoradiography. Distribution and subcellular localization were determined in human neutrophils, human epithelial carcinoma cells (A431) and in rat kidney tissue