Biolegend
Alexa Fluor® 647 anti-mouse IFN-γ Antibody
505814
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After stimulating CD8+ T cells with differential amounts of antigen, we used AF647 anti-mouse IFN-γ antibody to measure IFN-γ intracellular cytokine production, compared to an isotype control.
Flow Cytometry
Mouse CD8+ T cells
N/A
30 mins staining for surface receptors (CD8+) on ice (1:200). Washed 3x with FACS buffer. Fixed and permed with 2% PFA and 4% sucrose for 30 min at RT before permeabilizing with 0.5% saponin.
Incubated AF647 IFN-γ antibody in perm buffer (0.5% saponin in FACS buffer) for 30 min on ice before washing 2x with perm buffer and 1x with FACS buffer.
Flow cytometry
High levels of IFN-γ were detected in CD8+ T cells 48-72 hours after stimulation with anti-CD3/CD28. Approximately 70% were positive for IFN-γ compared to isotype control.
None
Successfully detects intracellular IFN-γ
Cannot really be used to quantify the total amount of IFN-γ produced.
Great antibody to use for measure percentage of cells that are producing intracellular IFN-γ.