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

Journal: European journal of biochemistry

Article Title: N-terminal and C-terminal plasma membrane anchoring modulate differently agonist-induced activation of cytosolic phospholipase A2.

doi: 10.1046/j.1432-1327.1999.00797.x

Figure Lengend Snippet: Fig. 2. Subcellular distribution of wild-type cPLA2, myc-cPLA2, Lck-cPLA2 and cPLA2-Ras in CHO-2B cells. Parental CHO-2B cells (A) and cells transiently transfected with either wild-type cPLA2 (B), myc-cPLA2 (C), Lck-cPLA2 (D) or cPLA2-Ras (E) constructs were stained with a polyclonal anti-cPLA2 and a FITC-conjugated second antibody (green). Nuclei were stained with propidium iodide (red). Immuno- fluorescent staining was visualized by confocal microscopy. Note the plasma membrane distribution of Lck-cPLA2 (D) and cPLA2-Ras (E) as opposed to the punctate cytoplasmic distribution of wild-type cPLA2 (B) and myc-cPLA2 (C). Immunofluorescence analysis was also performed on stable transfectants producing the various forms of cPLA2. The patterns of staining were similar to those of the transiently transfected cells. Bar, 10 mm.

Article Snippet: They were then incubated with fluorescein isothiocyanate (FITC)-conjugated donkey anti-(rabbit IgG) (diluted 1 : 100; Jackson ImmunoResearch Laboratories) for 1 h, washed and incubated for 10 min with 1 mg ́mL21 RNAse A.

Techniques: Transfection, Construct, Staining, Confocal Microscopy, Clinical Proteomics, Membrane, Immunofluorescence