Fluorescent phycobiliprotein conjugates for analyses of cells and molecules.

摘要: The synthesis of a novel class reagents for fluorescence analyses molecules and cells is reported. These compounds consist highly fluorescent phycobiliprotein conjugated to molecule having biological specificity. Phycoerythrin-immunoglobulin, phycoerythrin-protein A, phycoerythrin-avidin conjugates were prepared. bind specifically beads containing covalently attached target render them fluorescent. Femtomole (10(-15) mole) quantities phycoerythrin can be detected because the high extinction coefficient (epsilon M = 2.4 x 10(6) cm-1 M-1 10(5) daltons) quantum yield (Q 0.8) moiety. An important feature these that they emit in orange-red spectral region, where background less than at shorter wavelengths. Phycoerythrin are well-suited two-color flow cytofluorimetric employing single excitation line. distributions Leu antigens (also called OKT antigens) on surface T-lymphocytes analyzed using fluoresceinated antibody as green-fluorescent stain biotinylated counter-stained with red one. This one-laser analysis showed express Leu-3a Leu-3b or neither antigen. In contrast, Leu-2a (a marker suppressor cytotoxic T-cells) helper inducer mutually exclusive. studies show applied fluorescence-activated cell sorting analysis, microscopy, immunoassay.