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As a global market leader with numerous subsidiaries and distributors, Miltenyi Biotec is committed to providing our customers around the world with the highest quality products. In addition to direct selling in more than 20 countries in North America, Europe and Asia/Pacific, Miltenyi Biotec also provides support for our customers through an extensive distributor network covering dozens of additional countries.
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CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. |
A: | B: |
Before separation |
Figure 1CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. | Figure 1CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. |
CD4+CD25- T cells |
Figure 1CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. | Figure 1CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. |
Isolated CD4+CD25+ regulatory T cells |
Figure 1CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. | Figure 1CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. |
The general aim of my research was to test the ability of Regulatory T cells from drug treated mice to suppress the proliferation of effector T cells. I used this product in order to isolate the CD4+CD25+ regulatory T cell population from spleens and lymph nodes of control and drug-treated mice. I chose this kit because of the time and ease of use.
This Kit is very useful for isolating CD4+CD25+ Tregs from LN and Spleen, in order to culture Tregs, using them for RT PCR or for adoptive cell transfer experiments. I especially use it for performing in vivo Treg suppression assays, in which you transfer Tregs together with naive CD4+ T cells into immunodeficient RAG-/- mice. In this assay, the Tregs should prevent the colitogenic activity of the transferred naive CD4+ T cells.
CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. |
A: | B: |
Before separation |
Figure 1CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. | Figure 1CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. |
CD4+CD25- T cells |
Figure 1CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. | Figure 1CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. |
Isolated CD4+CD25+ regulatory T cells |
Figure 1CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. | Figure 1CD4 +CD25 + regulatory T cells were isolated from mouse spleen cell suspension by using the CD4 +CD25 + Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD25-PE and CD4‑FITC (A) or Anti-FoxP3-APC (B) and analyzed by flow cytometry using the MACSQuant ® Analyzer. Cell debris and dead cells were excluded from analysis based on scatter signals and propidium iodide fluorescence. |
The general aim of my research was to test the ability of Regulatory T cells from drug treated mice to suppress the proliferation of effector T cells. I used this product in order to isolate the CD4+CD25+ regulatory T cell population from spleens and lymph nodes of control and drug-treated mice. I chose this kit because of the time and ease of use.
This Kit is very useful for isolating CD4+CD25+ Tregs from LN and Spleen, in order to culture Tregs, using them for RT PCR or for adoptive cell transfer experiments. I especially use it for performing in vivo Treg suppression assays, in which you transfer Tregs together with naive CD4+ T cells into immunodeficient RAG-/- mice. In this assay, the Tregs should prevent the colitogenic activity of the transferred naive CD4+ T cells.
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