The following antibodies were used: allophycocyanin (APC)-H7–conjugated anti-CD3 (clone SK7), FITC-conjugated anti-CD4 (clone RPA-T4), APC-conjugated anti-CD4 (clone RPA-T4), Pacific blue (PB)–conjugated anti-CD4 (clone RPA-T4), APC-H7–conjugated anti-CD8 (clone SK1), PerCP-Cy5.5–conjugated anti-CD8 (clone SK1), PerCP-conjugated anti-CD69 (clone L78), PECy7-conjugated anti-CD279 (PD-1; clone EH12.1), APC-conjugated anti-TNF (MAbII), FITC-conjugated anti-CD25 (OX-39), PECy-conjugated anti-TNF (clone MabII; BD); energy coupled dye (ECD)–conjugated anti-CD3 (clone UCHT1), ECD-conjugated anti-CD45RA (clone 2H4), ECD-conjugated anti-CD4 (clone T4; Beckman Coulter); PECy5.5-conjugated anti-2B4 (CD244; clone C1.7), PE-conjugated anti-SLAM (CD150; clone A12), PB-conjugated anti-CD57 (clone HCD57), Alexa Fluor 647–conjugated anti-CD160 (clone BY55; BioLegend); EFluor 625NC-conjugated anti-CD8 (clone RPA-T8; eBioscience); and FITC-conjugated anti-CCR7 (clone 150503) and Alexa Fluor 700–conjugated anti–HLA-DR (clone LN3; R&D Systems).
Ecd conjugated anti cd3
Manufactured by Beckman Coulter
Sourced in United States
The ECD-conjugated anti-CD3 is a laboratory reagent designed for use in flow cytometry applications. It is a monoclonal antibody that recognizes the CD3 antigen, which is expressed on the surface of T cells. The ECD fluorochrome is conjugated to the anti-CD3 antibody, allowing for the detection and identification of T cells in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Pecy7 conjugated anti cd56, by BioLegend (2 mentions)
Pacific blue conjugated anti cd57, by BioLegend (2 mentions)
Pe conjugated nkg2c, by R&D Systems (2 mentions)
Fitc conjugated anti ccr7 clone 150503, by R&D Systems (1 mentions)
Percp cy5.5 conjugated anti cd8 clone sk1, by BD (1 mentions)
Apc conjugated anti tnf, by BD (1 mentions)
Apc h7 conjugated anti cd8, by BD (1 mentions)
Ldh activity assay kit, by Solarbio (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Sw1990, by American Type Culture Collection (1 mentions)
4 protocols using ecd conjugated anti cd3
1
Multicolor Flow Cytometry Panel
2
Adaptive NK Cells in CMV Reactivation
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Peripheral blood mononuclear cells (PBMCs) from HCT recipients were isolated from peripheral blood samples by density gradient centrifugation and analyzed by fluorescence-activated cell sorting (FACS) using an LSR II (BD Biosciences). PBMCs from recipients that reactivated CMV were collected at viral diagnosis, at 2, 4 and 8 weeks after antiviral therapy and at 6 months and 1 year post-transplant. For recipients that were CMV seronegative or were CMV seropositive without viral reactivation, PBMCs were collected at day 100, 6 months and 1 year post-transplant. The following fluorescently conjugated antibodies were used for phenotypic analysis: ECD-conjugated anti-CD3 (Beckman Coulter; IM2705U), PECy7-conjugated anti-CD56 (BioLegend; 318318), Pacific Blue-conjugated anti-CD57 (BioLegend; 322316) and PE-conjugated NKG2C (R&D Systems FAB138P-025). For statistical comparisons of adaptive NK cell percentages and absolute counts between RIC and MA recipients, unpaired, two-sided t-tests calculated using GraphPad were used. Error bars represent SEM. GraphPad was used to calculate R2 values and associated p values for the correlation between absolute monocyte and lymphocyte counts and adaptive NK cell expansion in 28 CMV seropositive recipients.
3
Adaptive NK Cells in CMV Reactivation
4
Evaluating T-cell Activation in Pancreatic Cancer
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Human pancreatic cancer cell lines (SW1990) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).
Cell transfection was performed using Lipofectamine 2000 (11668030, Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA). To examine T-cell activation, peripheral blood mononuclear cells (PBMCs) and tumor cells were cocultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS; 04-001-1A; Biological Industries, Göttingen, Germany) and 0.01% penicillin/streptomycin. The cells were then incubated with KrO-conjugated anti-CD45 (B36294; Beckman Coulter, Brea, CA, USA), ECD-conjugated anti-CD3 (A07748; Beckman Coulter), A700-conjugated anti-CD8 (737659; Beckman Coulter), APC-conjugated anti-CD69 (A80711; Beckman Coulter), and PE-conjugated anti-programmed cell death protein 1 (PD-1; B30634; Beckman Coulter) at 25 ℃ for 30 min and analyzed via flow cytometry, data analyzed using FlowJo software (BD Biosciences).
After coculture, the CD8+ T cells were isolated from PBMCs by CD8+ T Cell Isolation Kit, human (130-094-156, Miltenyi Biotec, German) and cocultured again with SW1990 cells. The cytotoxicity of CD8+ T cells was assessed using a lactate dehydrogenase (LDH) activity assay kit (BC0685; Solarbio, Beijing, China).
Cell transfection was performed using Lipofectamine 2000 (11668030, Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA). To examine T-cell activation, peripheral blood mononuclear cells (PBMCs) and tumor cells were cocultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS; 04-001-1A; Biological Industries, Göttingen, Germany) and 0.01% penicillin/streptomycin. The cells were then incubated with KrO-conjugated anti-CD45 (B36294; Beckman Coulter, Brea, CA, USA), ECD-conjugated anti-CD3 (A07748; Beckman Coulter), A700-conjugated anti-CD8 (737659; Beckman Coulter), APC-conjugated anti-CD69 (A80711; Beckman Coulter), and PE-conjugated anti-programmed cell death protein 1 (PD-1; B30634; Beckman Coulter) at 25 ℃ for 30 min and analyzed via flow cytometry, data analyzed using FlowJo software (BD Biosciences).
After coculture, the CD8+ T cells were isolated from PBMCs by CD8+ T Cell Isolation Kit, human (130-094-156, Miltenyi Biotec, German) and cocultured again with SW1990 cells. The cytotoxicity of CD8+ T cells was assessed using a lactate dehydrogenase (LDH) activity assay kit (BC0685; Solarbio, Beijing, China).
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