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PerCP/Cy5.5 anti-mouse IFN-γ Antibody
505822
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Mouse transgenic OT-1 cells were stimulated with their cognate peptide, OVA, for 3 days. On the last day, cells were re-stimulated with PMA/ionomycin and treated with Brefeldin A to allow accumulation of protein in the cytoplasm. Following this, the cells were fixed in 1% formalin and then stained with this anti-mouse IFN-gamma antibody. Intracellular staining of cytokines is a difficult process and takes a lot of standardizing. However, with this antibody the results were obtained easily.
Flow Cytometry
Purified mouse CD8 T cells
1:200 dilution for 1 hour on ice
NA
Flow cytometry on PERCP channel
The OVA peptide stimulated cells show increase in IFN-gamma expression, which is expected. The fluorescence shift is not so great, but that maybe attributed to lower expression levels.
None
It works
Lots of standardization is required
A great antibody to start learning intracellular staining of cytokines.