The pooled PBMCs and lymph node cells from the healthy donors and the lung cancer patients were stained for flow cytometry. The following panel of mouse anti-human mAbs, all purchased from BD Biosciences (San Jose, CA, USA) or eBioscience (San Diego, CA, USA), was used: anti-human CD3-APC.cy7 (BD, 557832, SK7), anti-human CD4-Percp.cy5.5 (BD, 560650, RPA-T4), anti-human CD45RA-FITC (eBioscience, 11-0458-42, HI100), and anti-human CCR7-PE.cy7 (BD, 557648, 150503). The cell data were acquired using a 10-laser Gallios (Beckman Coulter Inc., Brea, CA, USA) analytical flow cytometer. Unstained and single fluorochrome-stained cells were used as controls to provide accurate compensation and data analysis. The results were analyzed with Kaluza software.
Anti human cd3 apc cy7
Manufactured by BD
Sourced in United States
Anti-human CD3-APC-Cy7 is a fluorescently-labeled antibody that binds to the CD3 antigen on human T cells. It is designed for use in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti human cd4 percp cy5, by BD (2 mentions)
Anti human cd95 pe cy7, by BD (2 mentions)
Anti human cd122 pe, by BD (2 mentions)
Lsrfortessa analytical flow cytometer, by BD (2 mentions)
Facscanto system, by BD (1 mentions)
Pe anti human cd25, by BioLegend (1 mentions)
Anti human cd4 bv510, by BD (1 mentions)
Anti human cd8 pe cy7, by BD (1 mentions)
Apc anti human cd8, by BioLegend (1 mentions)
Anti human cd49f fitc, by BioLegend (1 mentions)
Anti human cd19 percp cy5, by BD (1 mentions)
Anti human cd14 pecf594, by BD (1 mentions)
Version 10, by FlowJo (1 mentions)
Rpa t8, by Thermo Fisher Scientific (1 mentions)
Rpa t4, by BD (1 mentions)
αcd28, by BioLegend (1 mentions)
Anti human cd4 percp, by BD (1 mentions)
Cxcr3 apc, by BD (1 mentions)
Cd4 bb515, by BD (1 mentions)
Horizon fixable viability stain 700, by BD (1 mentions)
6 protocols using anti human cd3 apc cy7
1
Multiparameter Flow Cytometry for Immune Cell Analysis
2
Multiparameter Flow Cytometry Immunophenotyping
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The flow cytometry antibodies used in this study were as follows: anti-human CD3-APC-Cy7 (BD, 557832); anti-human CD4-BV510 (BD, 562970); anti-human CD8-PE-Cy7 (BD, 557746); anti-human CD45RA-BV605 (BD, 562886); anti-human CCR7-APC-R700 (BD, 565867); anti-human-CD8-APC (BioLegend, 300912); anti-human-CD25-PE (BioLegend, 302605); anti-human CD26-PE (BioLegend, 302705); anti-human CD49f-FITC (BioLegend, 313605); and the fluorescent dye 7-AAD (BioLegend, 420404). The cells were incubated with 1% Fc receptor blocker for 10 minutes and incubated with antibodies for 30 minutes. Flow cytometry analyses were conducted using the BD FACS Canto system. The data were analyzed with FlowJo software.
3
Phenotyping Extracellular Vesicles by Flow Cytometry
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Our specimen preparation protocol was modified from the methods suggested by Headland et al. (13 (link)). The cellular origin of plasma derived EVs were identified by the following CD markers; CD235 for erythrocytes, CD41A for platelets, CD146 for endothelial cells, CD66b for neutrophils, CD14 for monocytes, CD19 for B lymphocytes, and CD3 for T lymphocytes. Five microliters of the isolated EVs samples were diluted with PBS to a total volume of 50 μL. The samples were incubated with an antibody cocktail, comprised of anti-human CD9-FITC (ImmunoTools, Friesoythe, Germany), anti-human CD235-PE (BD Pharmingen, NJ, US), anti-human CD41A-PE-Cy5 (BD Pharmingen, NJ, US), anti-human CD146-PE-Cy7 (BD Pharmingen, NJ, US), anti-human CD66b-PE (BD Pharmingen, NJ, US), anti-human CD14-PECF594 (BD Pharmingen, NJ, US), anti-human CD19-PERCP-Cy5.5 (BD Pharmingen, NJ, US), and anti-human CD3-APC-Cy7(BD Pharmingen, NJ, US), for 1 h in the dark at room temperature. Flow cytometry data acquisition was done immediately after the antibody incubation process.
4
Phenotypic Analysis of T Cells in Chronic Pancreatitis
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T lymphocytes from the patients with CP were stained for flow cytometry. The following panel of mouse anti-human monoclonal antibodies (mAbs) used were all purchased from BD Biosciences (Franklin Lakes, NJ, USA) or eBioscience (San Diego, CA, USA): anti-human CD3-APC.Cy7 (cat. 557832, SK7; BD Biosciences), anti-human CD4-Percp.Cy5.5 (cat. 560650, RPA-T4; BD Biosciences), anti-human CD8-Alexa Fluor 700 (cat. 56-0088-42, RPA-T8; eBioscience), anti-human CCR7-PE.eFluor610 (cat. 61-1979-42, 3D12; eBioscience), anti-human CD62L-APC (cat. 17-0629-42, DREG56; eBioscience), anti-human CD45RO-FITC (11-0457-42, UCHL1; eBioscience), anti-human CD95-PE.Cy7 (cat. 561633, DX2; BD Biosciences), anti-human CD103-APC (cat. 17-1037-41, Ber-ACT8; eBioscience), and anti-human CD122-PE (cat. 554525; BD Biosciences). Cellular data were obtained from a BD LSRFortessa analytical flow cytometer. Unstained and single fluorochrome-stained cells served as controls for accurate compensation and data analysis. Cells in each sample were counted, and the data were analyzed with FlowJo version 10 (FlowJo, Ashland, OR, USA).
5
Characterization of T-cell Subset Dynamics
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The TILs from lung cancer patients and HD-PBMCs from healthy donors were incubated in 24-well plates precoated with 1 µg/mL α-CD3 (cat. 317315; Biolegend) in complete X-VIVO 15 medium contained 1 µg/mL α-CD28 (cat. 302923; Biolegend) and 1000 UI/mL IL-2, with or without 7 µM TWS119 (cat. S1590; Selleckchem) for 9 days. TILs and HD-PBMCs cells were harvested on days 3, 5, 7, 9, and then stained for flow cytometry. The following panel of mouse anti-human mAbs, all purchased from BD Biosciences (San Jose, CA) or eBioscience (San Diego, CA), were used: anti-human CD3-APC.Cy7 (BD, 557832, SK7), anti-human CD4-Percp.Cy5.5 (BD, 560650, RPA-T4), anti-human CD45RA-FITC (11-0458-42, HI100; eBioscience), anti-human CCR7-AF700 (BD, 561143, 150503), anti-human CD62L-PE-CF594 (BD, 562301, Dreg-56), anti-human CD95-PE.Cy7 (BD, 561633, DX2), and anti-human CD122-PE (BD, 554525). The cell data were acquired using a BD LSRFortessa analytical flow cytometer. Unstained and single fluorochrome-stained cells were served as controls to provide accurate compensation and data analysis. Cells were recorded per sample and the data were analyzed with FlowJo (version 10).
6
Immunophenotyping of T cells from BAL and blood
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Dead cells were stained with BD Horizon™ Fixable Viability Stain 700. Moreover, the expression of markers on T cells from BAL fluid and blood was stained with antibodies, including anti‐human CD3‐APC/Cy7, CD25‐APC, CD4‐BB515, CXCR3‐APC, CCR4‐PE, CCR5‐APC, and CCR8‐PE (BD Biosciences). Cells were acquired with a flow cytometer (Beckman Coulter), and data were analyzed with FlowJo 10.
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