Large Scale IFN-γ Secretion Assay – Enrichment Kit ( #130-091-329 )

Large Scale IFN-γ Secretion Assay – Enrichment Kit

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.

Disclaimer

For availability in your country, please contact your local representative.
MicroBeads are for research use only, not for use in humans.

Further information

Expansion of antigen-specific cytokine-secreting T cells
[PDF; 275,4 KB]

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.

Restimulation of antigen-specific cells is easily performed <i>in vitro</i>. 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.

B: Upon labeling with the Cytokine Catch Reagent, cytokine-specific antibodies are attached to the surface of all leukocytes via binding to CD45.

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.

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.

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.

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.

E: Magnetic enrichment of the cytokine-secreting cells is achieved using the CliniMACS ^plus Instrument and specifically designed separation software.

Magnetic enrichment of the cytokine-secreting cells is achieved using the CliniMACS <sup>plus</sup> Instrument and specifically designed separation software.

F: (Optional): For flow cytometric analysis the cytokine-secreting cells can be stained with fluorochrome-conjugated Cytokine Detection antibodies.

(Optional): For flow cytometric analysis the cytokine-secreting cells can be stained with fluorochrome-conjugated Cytokine Detection antibodies.