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| Large Scale IFN-γ Secretion Assay - Enrichment Kit |
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| Description |
The Large Scale IFN-γ Secretion Assay - Enrichment Kit is suitable for the enrichment of human IFN-γ-secreting, antigen-specific T cells from leukapheresis harvests, PBMCs, or other leukocyte-containing single-cell preparations.
The Large Scale IFN-γ Secretion Assay Enrichment Kit is exclusively designed for research use. Nevertheless, it can be used in combination with the manual cell separation systems as well as the autoMACS™Separator, the autoMACS™ Pro Separator, and the CliniMACS® Plus Instrument.
Different from the standard Cytokine Secretion Assays, this assay uses a direct strategy for the magnetic labeling of the target cells. The MicroBeads are directly conjugated to a secondary IFN-γ-specific antibody (see figure 1). The kit includes Anti-IFN-γ-PE staining antibodies for flow cytometric analysis. |
| Applications |
| The Large Scale IFN-γ Secretion Assay - Enrichment Kit allows the concomitant enrichment of viable CD4- and CD8-positive IFN-γ secreting T cells, e.g., for the large-scale generation of antigen-specific cell lines or clones. Tedious subcloning is not necessary and both CD4+ and CD8+ T cell lines may easily be expanded. |
| Equipment |
| Please inquire about recommended device and accessories. |
| Disclaimer |
For availability in your country, please contact your local representative.
MicroBeads are for research use only, not for use in humans. |
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| Figure 1 |
| Principle of the MACS® Cytokine Secretion Assay Technology using direct magnetic labeling of the target cells. |
| A: Restimulation of antigen-specific cells is easily performed in vitro. Depending on the nature of the specific antigen, cells are incubated for 3-16 hours. As a result, the responding cells rapidly begin to secrete cytokines. |
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| B: Upon labeling with the Cytokine Catch Reagent, cytokine-specific antibodies are attached to the surface of all leukocytes via binding to CD45. |
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| C: Cells are allowed to secrete cytokines for a period of 45 minutes at 37 °C. The secreted IFN-γ is exclusively captured by the IFN-gamma Catch Matrix Reagent on the surface of the cytokine-secreting cells. |
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| D: The cells are then labeled with MACS MicroBeads conjugated to a second IFN-γ-specific antibody (IFN-gamma Enrichment Reagent). The cells are now ready for magnetic enrichment. |
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| E: Magnetic enrichment of the cytokine-secreting cells is achieved using the CliniMACS plus Instrument and specifically designed separation software. |
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| F: (Optional): For flow cytometric analysis the cytokine-secreting cells can be stained with fluorochrome-conjugated Cytokine Detection antibodies. |
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| Products |
| Large Scale IFN-γ Secretion Assay - Enrichment Kit, human |
for 1×109 total cells
Download data sheet
130-091-329
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| MACS References |
| 1. Campbell J.D. (2003) Methods 31(2): 150-159.[8475] |
| 2. Feuchtinger T. et al. (2004) Exp. Hematol. 32(3): 282-289.[4334] |
| 3. Rauser G. et al. (2004) Blood 103(9): 3565-3572.[5341] |
| 4. Feuchtinger et al. (2006) Br. J. Haematol. 134: 64-76.[9717] |
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