Anti mouse cd8a percp

2×10⁶ PMBC were stained with PE H-2K^d AMQMLKETI MHC-I dextramer (Immudex) for 10 minutes in PBS with 5% FBS at room temperature. Anti-mouse CD3 Pac Blue, Anti-mouse CD8a PerCP, Anti-mouse CD127 APC (BD Bioscience) and Anti-mouse CD62L FITC (eBioscience) antibodies were added and incubated for 20 minutes at 4°C. Cells were washed twice with PBS 5% FBS then run immediately on a BD Biosciences LSR-Fortessa cell analyzer. CD3+CD8+ cells were analyzed for dextramer staining. The memory phenotype of dextramer positive CD8+ T cells was determined by analyzing the expression of CD62L and CD127 [47] (link). Central memory T cells were characterized as CD127+ CD62L+, effector memory CD127+ CD62L-, effector cells CD127- CD62L-, and transitional memory cells CD127- CD62L+.

Antibodies used include: anti-mouse CD45 APCCY7 (Clone 30-F11 Biolegend Cat #103116) and anti-mouse CD45 Eflour 450 (Clone 30-F11 Invitrogen Cat#48-0451-82); anti-mouse LY6G (Clone 1A8; Biolegend Cat#127623); anti-Mouse CD3 Pacific Blue (Clone 145-2C11 Biolegend Cat#100334); anti-Mouse CD3e, PerCP-Cy5.5 (Clone: 145-2C11, eBioscience, Cat#: 45-0031-82); anti-Mouse CD8a PerCP (Clone Ly-2 BDBiosciences Cat#M037858); anti-Mouse CD4, FITC (Clone: GK1.5; eBioscience, Cat#: 25-0041-82); anti-mouse CD19 PE (Clone eBio1D3 ThermoFisher Cat# 12-0193-83); anti-mouse CD11c APC (Clone N418; Biolegen Cat #117310); anti-mouse CD11b FITC (Clone M1/70.15 Invitrogen Cat #RM2801); anti-mouse CD25 PE (Clone 3C7 Biolegend Cat#101904); anti-mouse CD44 PE (Clone 1M7; Biolegend Cat# 10,008); anti-mouse CD62L APC (Clone Mel-14 Biolegend Cat# 104411).