Journal: The Journal of biological chemistry

Article Title: Rapid redistribution of CD20 to a low density detergent-insoluble membrane compartment.

doi: 10.1074/jbc.273.1.344

Figure Lengend Snippet: FIG. 1. FACScan profiles of CD20 mAb binding to Raji cells. Indirect immunofluorescence was performed as described under “Ex- perimental Procedures.” Solid profile represents the binding of the fluorescein isothiocyanate-labeled secondary antibody. Open profiles show binding of the primary antibodies 1F5, 2H7, and B1 as indicated.

Article Snippet: Bands were visualized using Kodak X-OMAT film (Eastman Kodak Co.) Immunofluorescence—Cells (2 3 105) were suspended and incubated in 100 ml of RPMI 1640 medium, 10% fetal bovine serum for 15 min at 37 °C with 2H7, 1F5, B1, or isotype-matched control mAb, washed once, and resuspended for 15 min with 100 ml of 1/100 dilution of goat anti-mouse IgG conjugated to fluorescein isothiocyanate (Southern Biotechnology Associates, Inc.).

Techniques: Binding Assay, Immunofluorescence, Labeling

Journal: The Journal of biological chemistry

Article Title: Rapid redistribution of CD20 to a low density detergent-insoluble membrane compartment.

doi: 10.1074/jbc.273.1.344

Figure Lengend Snippet: FIG. 5. 2H7-induced CD20 redistribution to the Triton-insolu- ble fraction is rapid and does not involve internalization. A, CD20 immunoblot. Cells were treated with 2H7 mAb for the times indicated and lysed immediately with 2 3 lysis buffer. Each lane con- tains Triton-insoluble material from 2 3 105 cells. B, indirect immuno- fluorescence. Solid profile represents the binding of the fluorescein isothiocyanate-labeled secondary antibody. Open profiles, cells were incubated for 15 min at 37 °C with 2H7 and then washed. Fluorescein isothiocyanate-conjugated secondary antibody was either added im- mediately (solid line) or after a further 45-min incubation at 37 °C (dashed line).

Article Snippet: Bands were visualized using Kodak X-OMAT film (Eastman Kodak Co.) Immunofluorescence—Cells (2 3 105) were suspended and incubated in 100 ml of RPMI 1640 medium, 10% fetal bovine serum for 15 min at 37 °C with 2H7, 1F5, B1, or isotype-matched control mAb, washed once, and resuspended for 15 min with 100 ml of 1/100 dilution of goat anti-mouse IgG conjugated to fluorescein isothiocyanate (Southern Biotechnology Associates, Inc.).

Techniques: Western Blot, Lysis, Fluorescence, Binding Assay, Labeling, Incubation