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CD14+ monocytes were isolated with CD14 MicroBeads, human, from peripheral blood mononuclear cells (PBMCs) and cultured in the presence of GM-CSF and interleukin 4 (IL-4) for 6 days into inmature monocyte-derived dendritic cells (Mo-DCs). For generation of mature Mo-DCs, cells were cultured for another 24 hours in the presence of TNF-α, IL-6, and IL-1β. Mo-DCs were then stained with CD80 antibodies or with the corresopnding isotype control antibodies (left peak) and analyzed by flow cytometry using the MACSQuant® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.
Other clones | Overlap in epitope recognition with 2D10 |
---|---|
REA661 | ++ |
QA18A16 | ++ |
W17149D | ++ |
L307.4 | + |
Cells were incubated with an excess of purified unconjugated CD80 (2D10) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker. |
Clone | 2D10 |
---|---|
Clonality | monoclonal |
Isotype | mouse IgG1κ |
Isotype control | Isotype Control Antibody, mouse IgG1 |
Host | mouse |
Type of antibody | Primary antibodies |
Species | human, non-human primate |
Cross-reactivity | rhesus monkey ( Macaca mulatta) |
Antigen | CD80 |
Alternative names of antigen | B7, B7-1, BB1, CD28LG, CD28LG1, LAB7 |
Molecular mass of antigen [kDa] | 29 |
Distribution of antigen | B cells, dendritic cells, macrophages, monocytes, Langerhans cells, T cells |
Entrez Gene ID | 941 |
RRID | AB_2733839, AB_2801992, AB_2801971, AB_2857834, AB_2857831, AB_2659263, AB_2733838 |
Other clones | Overlap in epitope recognition with 2D10 |
---|---|
REA661 | ++ |
QA18A16 | ++ |
W17149D | ++ |
L307.4 | + |
Cells were incubated with an excess of purified unconjugated CD80 (2D10) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker. |
Clone | 2D10 |
---|---|
Clonality | monoclonal |
Isotype | mouse IgG1κ |
Isotype control | Isotype Control Antibody, mouse IgG1 |
Host | mouse |
Type of antibody | Primary antibodies |
Species | human, non-human primate |
Cross-reactivity | rhesus monkey ( Macaca mulatta) |
Antigen | CD80 |
Alternative names of antigen | B7, B7-1, BB1, CD28LG, CD28LG1, LAB7 |
Molecular mass of antigen [kDa] | 29 |
Distribution of antigen | B cells, dendritic cells, macrophages, monocytes, Langerhans cells, T cells |
Entrez Gene ID | 941 |
RRID | AB_2733839, AB_2801992, AB_2801971, AB_2857834, AB_2857831, AB_2659263, AB_2733838 |
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