Table 2.

Flow cytometry antibodies and reagents to measure intracellular expression of TNF and IFNɣ, and Granzyme B in CD8+ T cells per 250,000 cocktail-stimulated T cells

Stain	Stain dilution
Intracellular Panel 1: Cocktail 1 - Cell Stimulation and Protein secretion inhibitor Cocktail
Viability Dye Fixable Aqua	1 in 2,000 in PBS
CD8 or CD3 in APC-Cy7	1 in 100 in FACS Buffer
IFNɣ in APC	1 in 50 in 1× Perm Buffer
TNF in PE	1 in 50 in 1× Perm Buffer
Isotype control for Intracellular Panel 1
Viability Dye Fixable Aqua	1 in 2,000 in PBS
CD8 or CD3 in APC-Cy7	1 in 100 in FACS Buffer
APC Mouse IgG1	1 in 50 in 1× Perm Buffer
PE Mouse IgG1	1 in 50 in 1× Perm Buffer
Gating control for Intracellular Panel 1, treated with Cocktail 2 - Protein secretion inhibitor Cocktail
Viability Dye Fixable Aqua	1 in 2,000 in PBS
CD8 or CD3 in APC-Cy7	1 in 100 in FACS Buffer
IFNɣ in APC	1 in 50 in 1× Perm Buffer
TNF in PE	1 in 50 in 1× Perm Buffer
Intracellular Panel 2: Cocktail 2 - Protein secretion inhibitor Cocktail
Viability Dye Fixable Aqua	1 in 2,000 in PBS
CD8 or CD3 in APC-Cy7	1 in 100 in FACS Buffer
Granzyme B in FITC	1 in 50 in 1× Perm Buffer
Isotype control for Intracellular Panel 2
Viability Dye Fixable Aqua	1 in 2,000 in PBS
CD8 or CD3 in APC-Cy7	1 in 100 in FACS Buffer
FITC Mouse IgG1	1 in 50 in 1× Perm Buffer