A dialysis technique for preparing fluorescent antibody

Binding of Soluble Immune Complexes to Human Lymphoblastoid Cells

“…(Miles Laboratories, Inc., Kankakee, Ill.) on a DEAE-52 cellulose column. IgG (10 mg/ml) was conjugated with fluorescein isothiocyanate (FITC) as in (11) and was aggregated subsequently by heating at 63°C for 20 min. Soluble fluorescein-conjugated aggregated human gamma globulin (FITC AHG), prepared after centrifugation of the aggregates at 1,500 g for 30 min., was used in a concentration of 2 mg/ml of phosphate-buffered saline (PBS), pH 8.0.…”

“…Antisera were conjugated with FITC according to the method of Clark and Shepard (11). In a typical experiment, 5 × 108 cells in 50t~l of MEM without Ca ++ and with 100 ttg of isolated human C3 or C3b or 50 ttl of human, mouse, rabbit or guinea pig serum (freshly obtained or previously stored at -70°C) were incubated with gentle agitation at 37°C for 30 min.…”

Specificities and binding properties of 2 monoclonal antibodies against carcinoma cells of the human urinary bladder

“…For IFL, the slides were washed and incubated for an additional 30min with rabbit F(ab')2 anti-mouse immunoglobulin (Ig) conjugated to fluorescein isothiocyanate (Clark & Shepard, 1963), washed again and mounted in 50% glycerol in PBS prior to examination. For immunoperoxidase staining, the slides were incubated with sheep F(ab')2 anti-mouse Immunoprecipitation and SDS-PAGE analysis of target components Immunoprecipitation was performed essentially as described earlier by Paulie et al (1984).…”

Neuraminidase sensitive antigenic determinants of plasma cell tumor membrane glycoproteins

“…After repeated washings with complete culture medium, cells were stained for 30 min at 0°C with the immunoglobulin fraction of a goat anti-rabbit L chain antiserum, FITC labelled [24]. Samples were examined with a Leitz Ortholux fluorescence microscope equipped with incident light and immersion objectives.…”