FluoroSpot Plus: Mouse IFN‑γ/IL‑2/TNF‑α
FluoroSpot Plus: Mouse IFN‑γ/IL‑2/TNF‑α
Components
| Plate | Pre-coated FluoroSpot plate (mAbs AN18, 1A12, and MT1C8/23C9) |
| Detection mAbs | Anti-IFN-γ mAb (R4-6A2), BAM |
| Anti-IL-2 mAb (5H4), biotin | |
| Anti-TNF-α mAb (MT11B10), WASP | |
| Fluorophore conjugates | Anti-BAM mAb, 490 |
| SA-550 | |
| Anti-WASP mAb, 640 | |
| Buffer/Solution | FluoroSpot enhancer |
| Co-stimulus | Anti-CD28 mAb |
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Performance
Mouse splenocytes (200,000 cells/well) were incubated overnight in the absence or presence of ConA and anti-CD28 mAb. The number of cells secreting IFN-γ, IL-2 and TNF-α was analyzed by FluoroSpot. Single cytokine secreting cells as well as double- and triple-secreting cells were determined. Spot center was used to identify spots from double- and triple-secreting cells, visualized in an image overlay of LED490, FITC (IFN-γ) and LED550, Cy3 (IL-2) and LED640, Cy5 (TNF-α). Spot analysis was made with Mabtech IRIS.
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Analyte information
IFN-γ
| Analyte description | Interferon-γ (IFN-γ) is the only type II interferon. This proinflammatory cytokine is secreted by activated T cells and NK cells. It activates macrophages and endothelial cells and regulates immune responses by affecting APCs, T cells, and B cells. Production of IFN-γ by helper T cells and cytotoxic T cells is a hallmark of the Th1-type phenotype. Thus, high-level production of IFN-γ is typically associated with effective host defense against intracellular pathogens. |
| Alternative names | Interferon-γ, Interferon-gamma, IFN-γ, IFN-gamma, IFN-g, IFNg, If2f, Ifg |
| Cell type | T cell, Tc, Th1, NK cell |
| Gene ID | 15978 |
IL-2
| Analyte description | Interleukin 2 (IL-2) is predominantly produced by activated T cells. IL-2 promotes the proliferation, differentiation, and survival of antigen-activated T cells and NK cells. After antigen stimulation, the cytokine also promotes the differentiation of T cells into effector and memory T cells. Therefore IL-2 plays an important role in the body's response to infection. IL-2 is also key to tolerance, as it promotes the differentiation of certain immature T cells into regulatory T cells in the thymus, ultimately preventing autoimmune diseases. |
| Alternative names | Interleukin-2, Interleukin 2, IL-2, IL2, T cell growth factor, Il-2 |
| Cell type | T cell, Th1, Th2, Th17, Tfh |
| Gene ID | 16183 |
TNF-α
| Analyte description | Tumor necrosis factor (TNF), also known as TNF-α, is produced by many different cell types, e.g., monocytes, macrophages, T cells, and B cells. Among the many effects of TNF-α are protection against bacterial infection, cell growth modulation, immune system regulation, and involvement in septic shock. |
| Alternative names | Tumor necrosis factor-α, TNF-α, TNF-alpha, TNF-a, TNFa, Tumor necrosis factor-alpha, TNF, DIF, TNFSF2, TNFalphaa, Tnfsf1a, Tnlg1f, Tnf |
| Cell type | T cell, Tc, Th1, Th2, Th17, Tfh, Monocyte/MΦ |
| Gene ID | 21926 |