Time-resolved immunofluorometric detection of antigens separated by high-performance liquid chromatography and coated to polystyrene
Diamandis, E.P.; Christopoulos, T.K.
Biotechniques 10(5): 646-648
1991
ISSN/ISBN: 0736-6205 PMID: 1716927 Document Number: 372673
We use high-performance liquid chromatography with fraction collection to separate an antigen of interest. The antigen is then immobilized on polystyrene microtiter wells and detected with a specific antibody, followed by a second biotinylated antibody and streptavidin labeled with a fluorescent europium chelate. Fluorescence can be quantified with the use of time-resolved fluorescence. Antigen detectability down to 2-3 x 10(-17) mol was achieved. This method could be used as an alternative to Western blot in certain applications.