Conjugação e validação de controle isotípico IgG1-FITC para uso em citometria de fluxo

Abstract It was during the 1950's that the development of flow cytometrystarted, technology that allow to measure physiochemicalcharacteristics of cells or suspended particles in fluid. This technologyuses monoclonal antibodies labeled by fluorochromes as investigationtool in several analysis and needs isotype controls to define thenegative region (background). These controls are constituted byimmunoglobulins of the same isotype and fluorochrome from testantibodies, being fluorescein isothiocyanate (FITC) the mostfluorescent marker used in antibody conjugations. The isotypecontrols have the function to define the unspecific fluorescence(negative cells) and the fluorescent regions (positive cells). In thisstudy was selected monoclonal antibody (mAb) against canineerythrocyte antigen, produced in the Monoclonal Antibodies Tabela 3. Medidas descritivas de leucocitos animais (%) segundo especie animalMedidas DescritivasEspecieCaninaDemaisEspeciesNo de animais 52 6Valor minimo 0,00 0,001o Quartil 0,00 0,03Mediana 0,02 0,063o Quartil 0,13 0,13Valor maximo 2,51 0,41Media 0,16 0,11Desvio-padrao 0,42 0,15Resultado do testeestatistico1,47 (P>0,05)Conclusao estatistica: Especie canina = Bovina + EquinaTabela 4. Medidas descritivas de leucocitos humanos (%)segundo reagenteMedidas DescritivasReagente