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Isolation of lineage marker-negative cells from mouse bone marrow using the Direct Lineage Cell Depletion Kit, a MidiMACS™ Separator and an LS Column. Cells were fluorescently labeled with the Lineage Cell Detection Cocktail-Biotin and Anti-Biotin-APC and analyzed by flow cytometry using the MACSQuant ® Analyzer. In order to evaluate the LSK fraction, cells were additionally labeled with CD117-PE and Anti-SCA-1-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence. |
Before separation | After separation | LSK staining |
Lineage Cell Depletion Kit, mouseFigure 1Isolation of lineage marker-negative cells from mouse bone marrow using the Direct Lineage Cell Depletion Kit, a MidiMACS™ Separator and an LS Column. Cells were fluorescently labeled with the Lineage Cell Detection Cocktail-Biotin and Anti-Biotin-APC and analyzed by flow cytometry using the MACSQuant ® Analyzer. In order to evaluate the LSK fraction, cells were additionally labeled with CD117-PE and Anti-SCA-1-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence. | Lineage Cell Depletion Kit, mouseFigure 1Isolation of lineage marker-negative cells from mouse bone marrow using the Direct Lineage Cell Depletion Kit, a MidiMACS™ Separator and an LS Column. Cells were fluorescently labeled with the Lineage Cell Detection Cocktail-Biotin and Anti-Biotin-APC and analyzed by flow cytometry using the MACSQuant ® Analyzer. In order to evaluate the LSK fraction, cells were additionally labeled with CD117-PE and Anti-SCA-1-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence. | Lineage Cell Depletion Kit, mouseFigure 1Isolation of lineage marker-negative cells from mouse bone marrow using the Direct Lineage Cell Depletion Kit, a MidiMACS™ Separator and an LS Column. Cells were fluorescently labeled with the Lineage Cell Detection Cocktail-Biotin and Anti-Biotin-APC and analyzed by flow cytometry using the MACSQuant ® Analyzer. In order to evaluate the LSK fraction, cells were additionally labeled with CD117-PE and Anti-SCA-1-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence. |
Our lab has generated a mouse model that expresses three transgenic human host factors and can support HIV infection. We are interested in understanding protective immune responses that could protect against HIV infection in these mice. To understand the role that CD8 T cell responses might have against HIV infection, we reprogram their precursor cells at stem cell level by introducing TCRs that are known to have potent anti-HIV activity. We use this kit to deplete all lineage committed cells from bone marrow.
Isolation of lineage marker-negative cells from mouse bone marrow using the Direct Lineage Cell Depletion Kit, a MidiMACS™ Separator and an LS Column. Cells were fluorescently labeled with the Lineage Cell Detection Cocktail-Biotin and Anti-Biotin-APC and analyzed by flow cytometry using the MACSQuant ® Analyzer. In order to evaluate the LSK fraction, cells were additionally labeled with CD117-PE and Anti-SCA-1-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence. |
Before separation | After separation | LSK staining |
Lineage Cell Depletion Kit, mouseFigure 1Isolation of lineage marker-negative cells from mouse bone marrow using the Direct Lineage Cell Depletion Kit, a MidiMACS™ Separator and an LS Column. Cells were fluorescently labeled with the Lineage Cell Detection Cocktail-Biotin and Anti-Biotin-APC and analyzed by flow cytometry using the MACSQuant ® Analyzer. In order to evaluate the LSK fraction, cells were additionally labeled with CD117-PE and Anti-SCA-1-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence. | Lineage Cell Depletion Kit, mouseFigure 1Isolation of lineage marker-negative cells from mouse bone marrow using the Direct Lineage Cell Depletion Kit, a MidiMACS™ Separator and an LS Column. Cells were fluorescently labeled with the Lineage Cell Detection Cocktail-Biotin and Anti-Biotin-APC and analyzed by flow cytometry using the MACSQuant ® Analyzer. In order to evaluate the LSK fraction, cells were additionally labeled with CD117-PE and Anti-SCA-1-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence. | Lineage Cell Depletion Kit, mouseFigure 1Isolation of lineage marker-negative cells from mouse bone marrow using the Direct Lineage Cell Depletion Kit, a MidiMACS™ Separator and an LS Column. Cells were fluorescently labeled with the Lineage Cell Detection Cocktail-Biotin and Anti-Biotin-APC and analyzed by flow cytometry using the MACSQuant ® Analyzer. In order to evaluate the LSK fraction, cells were additionally labeled with CD117-PE and Anti-SCA-1-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence. |
Our lab has generated a mouse model that expresses three transgenic human host factors and can support HIV infection. We are interested in understanding protective immune responses that could protect against HIV infection in these mice. To understand the role that CD8 T cell responses might have against HIV infection, we reprogram their precursor cells at stem cell level by introducing TCRs that are known to have potent anti-HIV activity. We use this kit to deplete all lineage committed cells from bone marrow.
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