We performed flow cytometry analysis using the following antibodies: CD3-Percp-Cy5.5 (HIT3a, BD Bioscience), CD8-APC (RPA-T8, BD Bioscience), CD27-PE (M-T271, BD Bioscience), CD28-APC (CD28.2, BD Bioscience), CD45RO-PE (UCHL1, BD Bioscience), CD62L-FITC (DRGE-56, BD Bioscience), αvβ3-FITC (LM6090, EMD Millipore,), αvβ5-FITC (P1F6, EMD Millipore), and NRP-1-PE (AD5-17F6, Miltenyi Biotec GmbH). All samples tested were suspended in FACS buffer and stained with indicated antibodies for 30 min in 4 °C in darks, and then, washed twice, and resuspended in FACS buffer before analysis. For IFN-γ detection, BD™ Cytometric Bead Array (CBA) Human IFN-γ kit (BD Bioscience, USA) was used. Samples were analyzed with BD Accuri C6 (BD Bioscience).
Cd27 pe m t271
Manufactured by BD
Sourced in United States
CD27-PE (M-T271) is a monoclonal antibody labeled with the fluorescent dye phycoerythrin (PE). It is designed for the detection and analysis of CD27-expressing cells using flow cytometry. CD27 is a member of the tumor necrosis factor receptor superfamily and is expressed on various immune cell types.
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Lab products found in correlation
Anti igd ab, by Southern Biotech (2 mentions)
Cd19 allophycocyanin hib19, by BD (2 mentions)
Cd38 pecy hb7, by BD (2 mentions)
Igd fitc ia6 2, by BD (2 mentions)
Anti biotin microbead, by Miltenyi Biotec (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Fortessa, by BD (2 mentions)
B cell isolation kit 2, by Miltenyi Biotec (2 mentions)
Cd8 apc rpa t8, by BD (1 mentions)
Cd45ro pe uchl1, by BD (1 mentions)
Cd28 apc cd28.2, by BD (1 mentions)
Lm6090, by Merck Group (1 mentions)
Accuri c6, by BD (1 mentions)
Cd19 apc cy7 hib19, by BioLegend (1 mentions)
Live dead fixable dead cell stain kit, by Thermo Fisher Scientific (1 mentions)
Lsrfortessa, by BD (1 mentions)
Cd45 v500 hi30, by BD (1 mentions)
Cd8 fitc sk1, by BD (1 mentions)
Cd3 apc r700 ucht1, by BD (1 mentions)
Cd4 apc h7 rpat4, by BD (1 mentions)
5 protocols using cd27 pe m t271
1
Comprehensive Immune Cell Profiling
2
Isolation and Characterization of Human B Cells
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B cells were isolated from PBMCs of healthy blood donors (Blood Transfusion Center Solna, Stockholm, Sweden) according [54 (link)]. Briefly, B cells were isolated either through negative selection (B Cell Isolation Kit II; Miltenyi Biotec) or by positive selection using biotinylated anti-IgD Ab (Southern Biotech) and Anti-Biotin MicroBeads (Miltenyi Biotec). The purity and B cell composition of each donor were assessed by flow cytometry, staining for CD19-allophycocyanin (HIB19; BD), IgD-FITC (IA6–2; BD), CD38-PECy (HB7; BD), CD27-PE (M-T271; BD), and DAPI (5.7 μM; Sigma-Aldrich) using an LSR II or Fortessa (BD) and analyzed using FlowJo software (TreeStar).
3
Comprehensive Immune Cell Phenotyping
4
Longitudinal Immune Profiling in Humanized Mice
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Immunophenotyping was performed on peripheral blood samples collected longitudinally and at the study endpoint on whole blood and on mononuclear cells isolated from the tissues of humanized mice. Prior to antibody incubation, Ig-binding sites were blocked. The fluorochrome-conjugated antibodies included CD45-V500 (HI30, BD Biosciences, San Jose, USA), CD3-APC-R700 (UCHT1, BD Biosciences, San Jose, USA), CD4-APC-H7 (RPA-T4, BD Biosciences, San Jose, USA), CD8-FITC (SK1, BD Biosciences, San Jose, USA), CD19-PE-Cy7 (SJ25C1, BD Biosciences, San Jose, USA), CD27-PE (M-T271, BD Biosciences, San Jose, USA), CD38-APC (HB7, BD Biosciences, San Jose, USA), CD45RA-Pacific Blue (F8-11-13, Bio-Rad, Hercules, USA), HLA-DR-PerCP (L243, BD Biosciences, San Jose, USA), mouse IgG1k-APC (MOPC-21, BD Biosciences, San Jose, USA), mouse IgG1k- Pacific Blue (MOPC-21, BD Biosciences, San Jose, USA), mouse IgG1k-PE (MOPC-21, BD Biosciences, San Jose, USA) and mouse IgG2ak-PerCP (X39, BD Biosciences, San Jose, USA). After antibody incubation, blood samples were lysed with t 1× BD FACS lysing solution (BD Biosciences, San Jose, USA). Samples were then analyzed on a BD LSRFortessa instrument (BD Biosciences, San Jose, USA).
5
Isolation and Characterization of B Cells
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