Journal: Glycobiology

Article Title: Galectin-loaded cells as a platform for the profiling of lectin specificity by fluorescent neoglycoconjugates: a case study on galectins-1 and -3 and the impact of assay setting.

doi: 10.1093/glycob/cwn009

Figure Lengend Snippet: Fig. 1. Loading of Raji cell surfaces with galectin-1 (A) and galectin-3 (B). Flow cytometric analysis of cells probed with the corresponding antibody preparations followed by application of secondary antibodies as described in Materials and methods. The logs of fluorescence intensity are plotted against cell number. The gray area defines background fluorescence of processed galectin-free cells (negative control). The number given for the black curve represents the percentage of cells reactive with the antibody, the standard for reactivity of the cell surface for galectins.

Article Snippet: Loading of Raji cell surfaces with galectins Raji cells (human B-lymphocyte origin, ATCC no. CCL86) were cultured in RPMI-1640 medium supplemented with 10% fetal calf serum and 2 mM glutamine at 37◦C in an atmosphere of 5% CO2.

Techniques: Negative Control

Journal: Glycobiology

Article Title: Galectin-loaded cells as a platform for the profiling of lectin specificity by fluorescent neoglycoconjugates: a case study on galectins-1 and -3 and the impact of assay setting.

doi: 10.1093/glycob/cwn009

Figure Lengend Snippet: Fig. 2. Probing the galectin-1-loaded Raji cells with fluorescent neoglycoconjugates in flow cytometry. The logs of fluorescence intensity are plotted against the cell number. The gray area defines background fluorescence of the galectin-free cells incubated with the probe. The numbers given for each black curve obtained for the series of glycocompounds (see name in the top part of each panel) represent the percentage of cells reactive with the fluorescent neoglycoconjugates, the measure for comparing galectin reactivity.

Article Snippet: Loading of Raji cell surfaces with galectins Raji cells (human B-lymphocyte origin, ATCC no. CCL86) were cultured in RPMI-1640 medium supplemented with 10% fetal calf serum and 2 mM glutamine at 37◦C in an atmosphere of 5% CO2.

Techniques: Cytometry, Incubation

Journal: Glycobiology

Article Title: Galectin-loaded cells as a platform for the profiling of lectin specificity by fluorescent neoglycoconjugates: a case study on galectins-1 and -3 and the impact of assay setting.

doi: 10.1093/glycob/cwn009

Figure Lengend Snippet: Fig. 3. Probing the galectin-3-loaded Raji cells with fluorescent neoglycoconjugates in flow cytometry. The logs of fluorescence intensity are plotted against the cell number. The gray area defines background fluorescence of galectin-free cells incubated with the probe. The numbers given for each black curve obtained for the series of glycocompounds (see name in the top part of each panel) represent the percentage of cells reactive with the fluorescent neoglycoconjugates, the measure for comparing galectin reactivity.

Article Snippet: Loading of Raji cell surfaces with galectins Raji cells (human B-lymphocyte origin, ATCC no. CCL86) were cultured in RPMI-1640 medium supplemented with 10% fetal calf serum and 2 mM glutamine at 37◦C in an atmosphere of 5% CO2.

Techniques: Cytometry, Incubation

Journal: Glycobiology

Article Title: Galectin-loaded cells as a platform for the profiling of lectin specificity by fluorescent neoglycoconjugates: a case study on galectins-1 and -3 and the impact of assay setting.

doi: 10.1093/glycob/cwn009

Figure Lengend Snippet: Fig. 4. Probing the galectin-loaded Raji cells with fluorescent neoglycoconjugates after incubation of cells with the plant lectin PNA in flow cytometry. The logs of fluorescence intensity are plotted against cell number. Gray area defines fluorescence of cells without pretreatment, black curves represent the fluorescence distribution of cell populations pretreated with PNA. (A) Galectin-1 probed with LNnT-PAA-fluo; (B) galectin-3 probed with Galα3′LN-PAA-fluo.

Article Snippet: Loading of Raji cell surfaces with galectins Raji cells (human B-lymphocyte origin, ATCC no. CCL86) were cultured in RPMI-1640 medium supplemented with 10% fetal calf serum and 2 mM glutamine at 37◦C in an atmosphere of 5% CO2.

Techniques: Incubation, Cytometry

Journal: Glycobiology

Article Title: Galectin-loaded cells as a platform for the profiling of lectin specificity by fluorescent neoglycoconjugates: a case study on galectins-1 and -3 and the impact of assay setting.

doi: 10.1093/glycob/cwn009

Figure Lengend Snippet: Fig. 5. Flow cytometric analysis of binding of galectin-1- and galectin-3 to Raji cells after enzymatic treatment of cells. Cells were treated with sialidase followed by β-galactosidase, and stained with SNA, MAA-I, PNA (A) or with anti-galectin antibodies (B). Fluorescence increase was calculated as [(Fi/F0) × 100] −100, where Fi is the fluorescence intensity of cells after incubation with digoxigenin-labeled lectins (A) or antibodies (B) and F0 is the fluorescence intensity of cells stained with secondary antibodies (B).

Article Snippet: Loading of Raji cell surfaces with galectins Raji cells (human B-lymphocyte origin, ATCC no. CCL86) were cultured in RPMI-1640 medium supplemented with 10% fetal calf serum and 2 mM glutamine at 37◦C in an atmosphere of 5% CO2.

Techniques: Binding Assay, Staining, Fluorescence, Incubation, Labeling

Journal: Glycobiology

Article Title: Galectin-loaded cells as a platform for the profiling of lectin specificity by fluorescent neoglycoconjugates: a case study on galectins-1 and -3 and the impact of assay setting.

doi: 10.1093/glycob/cwn009

Figure Lengend Snippet: Fig. 7. Probing galectin-1- and galectin-3-loaded Raji cells after enzymatic degalactosylation of the cells with β-galactosidase. Staining with DSA (A), anti-galectin-3 antibody (B), or probed with fluorescent LN-PAA (C, D). Fluorescence increase was calculated as [(Fi/F0) × 100] −100, where Fi is the fluorescence intensity of galectin-loaded cells after incubation with the secondary antibody (A, B) or with the probe and F0 is the fluorescence intensity of galectin-free cells exposed to secondary antibody (A, B) or to LN-PAA-fluo (C, D). (A) Data for galectin-3; similar results were obtained in the case of galectin-1.

Article Snippet: Loading of Raji cell surfaces with galectins Raji cells (human B-lymphocyte origin, ATCC no. CCL86) were cultured in RPMI-1640 medium supplemented with 10% fetal calf serum and 2 mM glutamine at 37◦C in an atmosphere of 5% CO2.

Techniques: Staining, Fluorescence, Incubation