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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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Human PBMCs from a SARS-CoV-2-reactive donor were incubated for 6 hours with a mix of the SARS-CoV-2 PepTivators ® Prot_S, Prot_M, and Prot_N or left unstimulated (negative control). Brefeldin A was added after 2 hours. Cells were then stained with the live/dead marker Viobility 405/452 Fixable Dye, fixed and permeabilized afterwards. Subsequently, cells were stained with the flow panel included in the kit. Cells were analyzed using a MACSQuant ® Analyzer 16. Doublets, debris, and dead cells as well as CD14 + and CD20 + cells were excluded. After pregating on CD3 as well as CD4 and CD8, respectively, activation marker and cytokine expression were assessed, e.g. CD154 and TNF-α for CD4 + T cells and TNF-α and IFN-γ for CD8 + T cells. |
A) CD4+ T cells |
Unstimulated | Stimulated |
Figure 1Human PBMCs from a SARS-CoV-2-reactive donor were incubated for 6 hours with a mix of the SARS-CoV-2 PepTivators ® Prot_S, Prot_M, and Prot_N or left unstimulated (negative control). Brefeldin A was added after 2 hours. Cells were then stained with the live/dead marker Viobility 405/452 Fixable Dye, fixed and permeabilized afterwards. Subsequently, cells were stained with the flow panel included in the kit. Cells were analyzed using a MACSQuant ® Analyzer 16. Doublets, debris, and dead cells as well as CD14 + and CD20 + cells were excluded. After pregating on CD3 as well as CD4 and CD8, respectively, activation marker and cytokine expression were assessed, e.g. CD154 and TNF-α for CD4 + T cells and TNF-α and IFN-γ for CD8 + T cells. | Figure 1Human PBMCs from a SARS-CoV-2-reactive donor were incubated for 6 hours with a mix of the SARS-CoV-2 PepTivators ® Prot_S, Prot_M, and Prot_N or left unstimulated (negative control). Brefeldin A was added after 2 hours. Cells were then stained with the live/dead marker Viobility 405/452 Fixable Dye, fixed and permeabilized afterwards. Subsequently, cells were stained with the flow panel included in the kit. Cells were analyzed using a MACSQuant ® Analyzer 16. Doublets, debris, and dead cells as well as CD14 + and CD20 + cells were excluded. After pregating on CD3 as well as CD4 and CD8, respectively, activation marker and cytokine expression were assessed, e.g. CD154 and TNF-α for CD4 + T cells and TNF-α and IFN-γ for CD8 + T cells. |
B) CD8+ T cells |
Unstimulated | Stimulated |
Figure 1Human PBMCs from a SARS-CoV-2-reactive donor were incubated for 6 hours with a mix of the SARS-CoV-2 PepTivators ® Prot_S, Prot_M, and Prot_N or left unstimulated (negative control). Brefeldin A was added after 2 hours. Cells were then stained with the live/dead marker Viobility 405/452 Fixable Dye, fixed and permeabilized afterwards. Subsequently, cells were stained with the flow panel included in the kit. Cells were analyzed using a MACSQuant ® Analyzer 16. Doublets, debris, and dead cells as well as CD14 + and CD20 + cells were excluded. After pregating on CD3 as well as CD4 and CD8, respectively, activation marker and cytokine expression were assessed, e.g. CD154 and TNF-α for CD4 + T cells and TNF-α and IFN-γ for CD8 + T cells. | Figure 1Human PBMCs from a SARS-CoV-2-reactive donor were incubated for 6 hours with a mix of the SARS-CoV-2 PepTivators ® Prot_S, Prot_M, and Prot_N or left unstimulated (negative control). Brefeldin A was added after 2 hours. Cells were then stained with the live/dead marker Viobility 405/452 Fixable Dye, fixed and permeabilized afterwards. Subsequently, cells were stained with the flow panel included in the kit. Cells were analyzed using a MACSQuant ® Analyzer 16. Doublets, debris, and dead cells as well as CD14 + and CD20 + cells were excluded. After pregating on CD3 as well as CD4 and CD8, respectively, activation marker and cytokine expression were assessed, e.g. CD154 and TNF-α for CD4 + T cells and TNF-α and IFN-γ for CD8 + T cells. |
Human PBMCs from a SARS-CoV-2-reactive donor were incubated for 6 hours with a mix of the SARS-CoV-2 PepTivators ® Prot_S, Prot_M, and Prot_N or left unstimulated (negative control). Brefeldin A was added after 2 hours. Cells were then stained with the live/dead marker Viobility 405/452 Fixable Dye, fixed and permeabilized afterwards. Subsequently, cells were stained with the flow panel included in the kit. Cells were analyzed using a MACSQuant ® Analyzer 16. Doublets, debris, and dead cells as well as CD14 + and CD20 + cells were excluded. After pregating on CD3 as well as CD4 and CD8, respectively, activation marker and cytokine expression were assessed, e.g. CD154 and TNF-α for CD4 + T cells and TNF-α and IFN-γ for CD8 + T cells. |
A) CD4+ T cells |
Unstimulated | Stimulated |
Figure 1Human PBMCs from a SARS-CoV-2-reactive donor were incubated for 6 hours with a mix of the SARS-CoV-2 PepTivators ® Prot_S, Prot_M, and Prot_N or left unstimulated (negative control). Brefeldin A was added after 2 hours. Cells were then stained with the live/dead marker Viobility 405/452 Fixable Dye, fixed and permeabilized afterwards. Subsequently, cells were stained with the flow panel included in the kit. Cells were analyzed using a MACSQuant ® Analyzer 16. Doublets, debris, and dead cells as well as CD14 + and CD20 + cells were excluded. After pregating on CD3 as well as CD4 and CD8, respectively, activation marker and cytokine expression were assessed, e.g. CD154 and TNF-α for CD4 + T cells and TNF-α and IFN-γ for CD8 + T cells. | Figure 1Human PBMCs from a SARS-CoV-2-reactive donor were incubated for 6 hours with a mix of the SARS-CoV-2 PepTivators ® Prot_S, Prot_M, and Prot_N or left unstimulated (negative control). Brefeldin A was added after 2 hours. Cells were then stained with the live/dead marker Viobility 405/452 Fixable Dye, fixed and permeabilized afterwards. Subsequently, cells were stained with the flow panel included in the kit. Cells were analyzed using a MACSQuant ® Analyzer 16. Doublets, debris, and dead cells as well as CD14 + and CD20 + cells were excluded. After pregating on CD3 as well as CD4 and CD8, respectively, activation marker and cytokine expression were assessed, e.g. CD154 and TNF-α for CD4 + T cells and TNF-α and IFN-γ for CD8 + T cells. |
B) CD8+ T cells |
Unstimulated | Stimulated |
Figure 1Human PBMCs from a SARS-CoV-2-reactive donor were incubated for 6 hours with a mix of the SARS-CoV-2 PepTivators ® Prot_S, Prot_M, and Prot_N or left unstimulated (negative control). Brefeldin A was added after 2 hours. Cells were then stained with the live/dead marker Viobility 405/452 Fixable Dye, fixed and permeabilized afterwards. Subsequently, cells were stained with the flow panel included in the kit. Cells were analyzed using a MACSQuant ® Analyzer 16. Doublets, debris, and dead cells as well as CD14 + and CD20 + cells were excluded. After pregating on CD3 as well as CD4 and CD8, respectively, activation marker and cytokine expression were assessed, e.g. CD154 and TNF-α for CD4 + T cells and TNF-α and IFN-γ for CD8 + T cells. | Figure 1Human PBMCs from a SARS-CoV-2-reactive donor were incubated for 6 hours with a mix of the SARS-CoV-2 PepTivators ® Prot_S, Prot_M, and Prot_N or left unstimulated (negative control). Brefeldin A was added after 2 hours. Cells were then stained with the live/dead marker Viobility 405/452 Fixable Dye, fixed and permeabilized afterwards. Subsequently, cells were stained with the flow panel included in the kit. Cells were analyzed using a MACSQuant ® Analyzer 16. Doublets, debris, and dead cells as well as CD14 + and CD20 + cells were excluded. After pregating on CD3 as well as CD4 and CD8, respectively, activation marker and cytokine expression were assessed, e.g. CD154 and TNF-α for CD4 + T cells and TNF-α and IFN-γ for CD8 + T cells. |
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