Bv421 anti cd45ra clone 5h9

CD3⁺CD8^– T lymphocytes were negatively selected from PBMCs and sorted on the BD FACS Aria III sorter into the following subsets: CD95^– naive T cells (CD45RA⁺CCR7⁺CD27⁺CD95^–), Tcm (CD45RA^–CCR7⁺CD27⁺), Ttm (CD45RA^–CCR7^–CD27⁺), and Tem cells (CD45RA^–CCR7^–CD27^–), as described in ref. 10 (link), using BV421 anti-CD45RA (clone 5H9, BD Biosciences), PE-Cy7 anti-CD27 (clone 0323, Invitrogen), APC anti-CD95 (clone DX2, BD Biosciences), and BB700 anti-CCR7 (clone 3D12, BD Biosciences). FlowJo v10.6 software was used for analysis.

CD3⁺CD8^– T lymphocytes were negatively selected from peripheral blood mononuclear cells (PBMCs) by an immunomagnetic bead–based protocol (Stem Cell Technologies, catalog 19052).
Purified T lymphocytes were then sorted by fluorescence activated cell sorting (BD FACS Aria III sorter) into the following subsets: CD3⁺CD8^– T_N (CD45RA⁺CCR7⁺CD27⁺), CD95^– T_N (CD45RA⁺CCR7⁺CD27⁺CD95^–), T_SCM (CD45RA⁺CCR7⁺CD27⁺CD95⁺), T_CM (CD45RA^–CCR7⁺CD27⁺), T_TM (CD45RA^–CCR7^–CD27⁺), T_EM (CD45RA^–CCR7^–CD27^–), and CD45RA_dim cells (Figure 1) using PE-Cy7 anti-CD27 (clone 0323, Invitrogen), BV421 anti-CD45RA (clone 5H9, BD Biosciences), BB700 anti-CCR7 (clone 3D12, BD Biosciences), and APC anti-CD95 (clone DX2, BD Biosciences). The following filters were used: 450/50 Violet for BV421, 780/40 Green for PeCy7, 710/50 Blue for CCR7, and 660/20 for APC (BD Biosciences). FlowJo v10.6 software was used for analysis.