A rapid objective immunofluorescence microassay. Application for detection of surface and intracellular antigens.
An indirect immunofluorescence microassay, which permits automated reading, has been employed for simple, rapid and objective detection of surface and intracellular antigens. Initially, the cells, spun in microplates, are fixed with glutaraldehyde (0.25% v/v in PBS). Following fixation, the cells can be stored at 4 degrees C for up to 2 weeks before being used in the immunofluorescence microassay. The fixed cells are then stained according to standard procedures using appropriate first and fluorescein-conjugated second antibodies. An automated and quantitative evaluation of the fluorescence intensity of the cell samples was achieved using the Titertek Fluoroskan II automatic reader. This microassay was shown to be suitable for the detection of the surface MAC1 antigen and intracellular v-myc protein in the GG2EE macrophage cell line.
Pitzurra
L
L
Department of Experimental Medicine and Biochemical Sciences, University of Perugia, Italy.
Blasi
E
E
Bartoli
A
A
Marconi
P
P
Bistoni
F
F
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000954:Antigens, Surface / Q000032:analysis*; D002460:Cell Line; D003593:Cytoplasm / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D016177:Macrophage-1 Antigen / Q000032:analysis; D008264:Macrophages / Q000276:immunology; D016274:Oncogene Protein p55(v-myc) / Q000032:analysis
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D016177:Macrophage-1 Antigen; D016274:Oncogene Protein p55(v-myc)