T lymphocytes execute and control immunological reactions with a repertoire of cytokines, cytotoxic substances, and other mediators. The quantitative and qualitative analysis of CD8
+
T cells specifically recognizing and reacting towards a defined antigen provide important information to understand their function in various immunological situations. Antigen-specific CD8
+
T cells can be identified and characterized by analyzing their effector function, e.g., production of cytokines.
The Rapid Cytokine Inspector (CD8 T Cell) Kit, human, has been developed for the fast and easy evaluation of cytokine expression in activated CD8
+
T cells

Data and images for Rapid Cytokine Inspector (CD8 T Cell) Kit, human

Figures

Figure 1

Human PBMCs were incubated with or without PepTivator CMV pp65 for 6 hours; brefeldin A was added after 2 hours. The cells were stained with the CD8 T Cell Detection Cocktail, Rapid Cytokine Inspector Anti-IFN-γ-APC, and Rapid Cytokine Inspector Anti- IL-2-PE. IFN-γ expression was analyzed for CD8
+
T cells in stimulated and unstimulated samples*.
A:
B:
Stimulated sample
Unstimulated control
View details

Figure 1

Human PBMCs were incubated with or without PepTivator CMV pp65 for 6 hours; brefeldin A was added after 2 hours. The cells were stained with the CD8 T Cell Detection Cocktail, Rapid Cytokine Inspector Anti-IFN-γ-APC, and Rapid Cytokine Inspector Anti- IL-2-PE. IFN-γ expression was analyzed for CD8
+
T cells in stimulated and unstimulated samples*.
View details

Figure 1

Human PBMCs were incubated with or without PepTivator CMV pp65 for 6 hours; brefeldin A was added after 2 hours. The cells were stained with the CD8 T Cell Detection Cocktail, Rapid Cytokine Inspector Anti-IFN-γ-APC, and Rapid Cytokine Inspector Anti- IL-2-PE. IFN-γ expression was analyzed for CD8
+
T cells in stimulated and unstimulated samples*.

Specifications for Rapid Cytokine Inspector (CD8 T Cell) Kit, human

Overview

T lymphocytes execute and control immunological reactions with a repertoire of cytokines, cytotoxic substances, and other mediators. The quantitative and qualitative analysis of CD8
+
T cells specifically recognizing and reacting towards a defined antigen provide important information to understand their function in various immunological situations. Antigen-specific CD8
+
T cells can be identified and characterized by analyzing their effector function, e.g., production of cytokines.
The Rapid Cytokine Inspector (CD8 T Cell) Kit, human, has been developed for the fast and easy evaluation of cytokine expression in activated CD8
+
T cells by intracellular staining. The kit contains an antibody cocktail for the identification of CD8
+
T cells and the exclusion of monocytes and B cells as well as brefeldin A and reagents for the fixation and permeabilization of cells after T cell stimulation. The kit is optimized for use in combination with Rapid Cytokine Inspector Anti-Cytokine antibodies of interest (not included in the kit).

Detailed product information

Detailed procedure

PBMCs are cultured with or without antigen for a total of 6 hours. Upon activation, the CD8
+
T cells start to produce cytokines. After 2 hours of cultivation, brefeldin A is added to the cells to inhibit transport of proteins to the cellular membrane. After stimulation, the culture supernatant is removed, and the CD8 T Cell Detection Cocktail is added together with Rapid Cytokine Inspector Anti-Cytokine antibodies of interest. Cell surface markers are stained during a short incubation step. Fixation and permeabilization of the cells is performed and enables subsequent intracellular staining of cytokines. The cells are washed and analyzed by flow cytometry. Washing steps after T cell stimulation, and before immunofluorescent staining, fixation, and permeabilization are not required.

Applications

● High-throughput multiparameter analysis of antigen-specific T cells.
● Rapid identification and enumeration of cytokine-producing activated antigen-specific CD8
+
T cells upon
in vitro
stimulation with the respective antigen or upon polyclonal restimulation.
● Immunomonitoring of antigen-specific T cells.

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