mouse anti rat cd4 r pe  (Bio-Rad)

Journal: Molecular Medicine

Article Title: Chlorogenic Acid Attenuates High Mobility Group Box 1 (HMGB1) and Enhances Host Defense Mechanisms in Murine Sepsis

doi: 10.2119/molmed.2012.00279

Figure Lengend Snippet: Percentage of various lymphocyte subpopulations.

Article Snippet: Fluorescent-labeled monoclonal antibodies, rat anti-mouse cluster of differentiation (CD) 8 (FITC) antibody and rat anti-mouse CD4 (R-PE) antibody (Invitrogen; Life Technologies), were used to quantify lymphocyte subpopulations present in the thymus, spleen and blood.

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: Early Murine T-lymphocyte Activation Is Accompanied by a Switch from N -Glycolyl- to N -Acetyl-neuraminic Acid and Generation of Ligands for Siglec-E *

doi: 10.1074/jbc.M111.243410

Figure Lengend Snippet: Activation of murine T-lymphocytes leads to up-regulation in the binding of siglec-E to cell surface proteins in a sialic acid-dependent manner. A , T-lymphocytes were activated with anti-CD3 and anti-CD28 for 24 h and then cultured for up to 4 days in the presence of IL-2. Cells harvested at each time point were incubated with fluorophore-conjugated CD4 ( left panel ) and CD8 antibodies ( right panel ) together with a pre-complex of FITC-conjugated goat anti-human Fc and siglec-Fc. Mean Fluorescence Intensity ( MFI ) of sialic acid-dependent binding by FACS is shown following subtraction of MFI values for sialidase-treated cells. B , for the detection of MAL and SNA binding, harvested cells corresponding to each time point were incubated with biotinylated lectin followed by a secondary incubation with FITC-conjugated streptavidin. C , splenocytes harvested at 24 h were untreated ( open histograms ) or pretreated with V. cholerae sialidase ( left panel ) or proteinase K ( right panel ) (shaded histograms) and labeled with siglec-E-Fc/anti-Fc-FITC complexes. Data are representative of at least three independent experiments.

Article Snippet: Cells were then incubated with pre-complexes for 30 min on ice together with 1/100 R-PE-conjugated rat anti-mouse CD4 and allophycocyanin-conjugated rat anti-mouse CD8 antibodies (Invitrogen).

Techniques: Activation Assay, Binding Assay, Cell Culture, Incubation, Fluorescence, Labeling

Journal: The Journal of Biological Chemistry

Article Title: Early Murine T-lymphocyte Activation Is Accompanied by a Switch from N -Glycolyl- to N -Acetyl-neuraminic Acid and Generation of Ligands for Siglec-E *

doi: 10.1074/jbc.M111.243410

Figure Lengend Snippet: T-lymphocyte activation is accompanied by alterations in the N -glycan profiles with increased proportion of NeuAc- to NeuGc-containing glycans. Permethylated N -glycans extracted from purified CD4+ and CD8+ T-lymphocytes, either resting (0 h), following 24 h activation, or following 24 h activation and 4 days cultivation in IL-2 were subjected to MS and MS/MS. The cartoon structures of selected peaks referred to in the text are illustrated.

Article Snippet: Cells were then incubated with pre-complexes for 30 min on ice together with 1/100 R-PE-conjugated rat anti-mouse CD4 and allophycocyanin-conjugated rat anti-mouse CD8 antibodies (Invitrogen).

Techniques: Activation Assay, Purification, Tandem Mass Spectroscopy

Journal: The Journal of Biological Chemistry

Article Title: Early Murine T-lymphocyte Activation Is Accompanied by a Switch from N -Glycolyl- to N -Acetyl-neuraminic Acid and Generation of Ligands for Siglec-E *

doi: 10.1074/jbc.M111.243410

Figure Lengend Snippet: Decrease in surface NeuGc immunoreactivity and CMAH gene expression following T-lymphocyte activation. Resting (0 h), activated (24 h), and activated + cultured (1 day and 4 days) T-lymphocytes were harvested and the binding of a chicken anti-NeuGc antibody to CD4+ ( A ) and CD8+ ( B ) cells was assessed by flow cytometry. Also included for comparison are data from siglec-E-Fc binding shown previously ( A ). Data are representative of at least three independent experiments. C , RNA was isolated from pooled T-lymphocytes at each time point and the expression of CMAH determined in triplicate relative to a normalized GAPDH control. Data show means ± S.D.

Article Snippet: Cells were then incubated with pre-complexes for 30 min on ice together with 1/100 R-PE-conjugated rat anti-mouse CD4 and allophycocyanin-conjugated rat anti-mouse CD8 antibodies (Invitrogen).

Techniques: Expressing, Activation Assay, Cell Culture, Binding Assay, Flow Cytometry, Isolation