GSM9891933: APECED_001_1753268_2_1_1; Homo sapiens; RNA-Seq
Blood was diluted 1:1 with PBS then added to SepMate-50 tubes filled with 15 mL of Ficoll-Paque PLUS (Cytiva, #17144003) and centrifuged at 1,200 g for 10 minutes. PBMCs and plasma layers were poured in a new tube, pelleted at 300 g for 10 minutes, and resuspended with 5 mL of ACK lysing buffer (Gibco, # A10492-01) for 3 minutes to eliminate red blood cells. PBS was added to 50 mL and PBMCs were pelleted at 300 g for 10 minutes. If necessary, the red blood cell lysis step was repeated. Portions of resuspended PBMCs were then used for single cell OMICs (scRNA/BCRseq) and high parameter flow cytometry. 10x 5prime NextGEM v2