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文献和实验The OP9-DL1 System: Generation of T-Lymphocytes from Embryonic or Hematopoietic Stem Cells In Vitro
al. 2006b ). MATERIALS Reagents Anti-CD24 monoclonal antibody (mAb) (J11d clone) (for Protocol 3) Use either culture supernatant that contains anti-CD24 mAb or purified anti-CD24 mAb (see Step 55). BDPharmLyse (red blood cell lysing
Combined Flow Cytometric Measurement of Two Cell-Surface Antigens and DNA-RNA Content
(Biodesign) in the presence of 100 ng/mL of surface-bound OKT3 (Ortho Diagnostics) (E-H ). (C and G ) Analysis of CD5+ CD8+ T cells; RNA/DNA dot-plots were gated on scatter as shown in row 1 (in A and E ) and R1 (in B ) or R2 (in F ), respectively. (D and H
个细胞,加入RPMI 1640培养基至总体积1ml。加入不同浓度的PMA(10ng、20ng和50ng终浓度)和500ng/ml Ionomycin共同刺激人外周血单个核细胞,同时加入0.7µl Golgistop,37 ℃ CO2孵箱刺激4h。收集单个核细胞,分为三部分,分别加入CD8-CYC、CD4-CYC、CD3-APC单抗各10μl,室温避光孵育30min。加入红细胞裂解液(FACS Lysing Solution)1ml裂解残余红细胞,2ml PBS洗涤细胞一次。流式
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