ebioscience/Thermo Fisher
IFN gamma Monoclonal Antibody (XMG1.2), PerCP-Cyanine5.5, eBioscience
45-7311-82
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Our lab studies the mTOR signaling pathway in T cell differentiation with an emphasis on both CD4 and CD8 T cells. We routinely conduct flow cytometry experiments to analyze the differentiation process into subsets based on transcription factor (TF) analysis and also cytokine production to assess function. In order to look at Th1 CD4 T cells, we analyze T cells for the expression of Tbet and also their capacity to produce IFNg, which are hallmark characteristics of Th1 cells. Using this antibody, we are able to detect IFNg secreting cells upon stimulation.
Flow Cytometry
Murine Splenocytes
1:500 at RT for 45 minutes (must be done as an intracellular stain)
FC blocking in FACS buffer (2% FBS, .2% NaAzide/PBS) before fixing/perming
None
Flow cytometer
By tracking proliferation using cell trace violet, we are able to identify proliferating T cells secreting IFNg. The signal is quite robust and are able to differentiate between producers versus non-producers.
N/A
The signal was very strong and was able to identify the cytokine producing population using a nice dilution.
Great reliable antibody to identify IFNg+ secreting T cells.