Fluorophore-conjugated murine antibodies were purchased from eBioscience, including antibodies specific for CD8a (53–6.7; catalog #48-0081-80), Thy1.1 (HIS 51; 25–0900-82), Tim3 (RMT3-23; 14–5870-81), Lag3 (ebioC9B7W; 12–2231-81), CD137 (17B5; 17-1370-80), KLRG1 (2F1; 175893–81), and CD25 (PC 61.5; 17–0251-81). Other antibodies to FasL (K10; 106805), vβ9 (MR10-2; 553201), CD69 (H1.2F3; 104502), CD127 (A7R34; 135013), CD80 (16-10A1; 553768), CD86 (GL-1; 105011), IL-2 (JES6-5H4; 503807), IFNγ (XMG1.2; 505809), TNFα (Mab11; 502913), Bax (6A7; 633801), and Bcl-2 (10C4; 633507) were purchased from Biolegend. 7AAD (A1310) and other antibodies were purchased from BD Biosciences, including Fas (Jo2; 563647), Thy1.2 (53–2.1; 561616), CD103 (M290; 557495), purified rat anti-mouse CD16/CD32 (2.4G2; 12–4875-80), PD-1 (J43; 11–9985-81), and Vβ8.1/8.2 (553185). Antibodies against pAKT (S473; D9E; 5315S) were purchased from Cell Signaling Technology. Anti-Eomes (DAN11 mag; 12–4875-80) and the LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (L34957) were purchased from ThermoFisher. For FasL detection, Panc1 cells were treated for 24 h with 3 µg/ml Batimastat (BB-94; ab146619) purchased from Abcam. Flow cytometry data were acquired on a BD FACS Canto II instrument or LSR II and analyzed with Flowjo v9 software.
Facsc
Manufactured by BD
Sourced in United States
The BD FACSC is a flow cytometry instrument used for the analysis and sorting of cells and particles in a liquid suspension. It is designed to measure and analyze the physical and fluorescent characteristics of individual cells or particles as they pass through a laser beam. The BD FACSC is a core tool in various research and clinical applications, providing researchers and clinicians with quantitative data on cell populations and their properties.
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Lab products found in correlation
Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions)
Purified rat anti mouse cd16 cd32, by BD (1 mentions)
Cd103, by BD (1 mentions)
Tnfα mab11, by BioLegend (1 mentions)
Vβ8 1 8, by BD (1 mentions)
Anti eomes dan11mag, by Thermo Fisher Scientific (1 mentions)
Batimastat bb 94, by Abcam (1 mentions)
Apoptosis detection kit, by Vazyme (1 mentions)
Fitc labeled annexin 5 pi apoptosis detection kit, by BD (1 mentions)
Facscalibur, by BD (1 mentions)
Annexin 5 fitc propidium iodide apoptosis detection kit, by Merck Group (1 mentions)
7 protocols using facsc
1
Fluorophore-conjugated antibodies for flow cytometry
2
Cell Cycle and Apoptosis Analysis
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For cell cycle analysis, drug-treated cells with 80% confluence were harvested and fixed with 70% ethanol. Then cells were taken for PI staining and cell cycle was analyzed using flow cytometry. For apoptosis analysis, cells were cultured in attachment, then trypsinized and stained with PI/Annexin V (Vazyme, Apoptosis Detection Kit). Data were collected and analyzed on a BD FACSC and using FACSD via software.
3
Cell Cycle and Apoptosis Analysis
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For cell cycle analysis, drug-treated cells with 80% confluence were harvested and fixed with 70% ethanol. Then cells were taken for PI staining and cell cycle was analyzed using flow cytometry. For apoptosis analysis, cells were cultured in attachment then trypsinized and stained with PI/Annexin V (Vazyme, Apoptosis Detection Kit). Data were collected and analyzed on a BD FACSC and using FACSD via software.
4
Apoptosis Analysis of Renal Cancer Cells
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786-O and A-498 cells were seeded into 24-well plates and treated with different concentrations of NCTD (0, 20, 40, or 80 μM) for 24 h. Cells were then harvested and washed with PBS, and stained with FITC-labeled Annexin V/PI Apoptosis Detection kit (BD Biosciences, CA, USA) at dark light for 20 min. Samples were subjected to flow cytometry (FACSCalibur, BD, USA), and data were collected and analyzed on a BD FACSC using FACSD software.
5
Cell Proliferation and Cell Cycle Analysis
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Cells were cultured in the 96‐well dishes at about density of 5000 cells/well and were detected at the 0, 1, 2 and 3 days. Each well was added with 10 μL CCK‐8 solution and incubated for 3 hours. The OD (optical density) was measured on spectrophotometer at 450 nm wavelength. Flow cytometry assay was utilized to study cell cycle. Cell was harvested and then washed in cold PBS and fixed by cold 70% ethanol overnight. Cell was incubated with PI (propidium iodide) solution on the ice for one half hour. Cell cycle was determined with flow cytometry (FACSC, BD).
6
Immune Cell Profiling in A. pleuropneumoniae Infection
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A. pleuropneumoniae-infected mice were sacrificed, and the lungs and spleens were collected and homogenized on ice. The homogenates were resuspended with PBS. After being washed twice, the cells were incubated with a fluorochrome-conjugated anti-cytokine antibody (APC-anti-CD3, FITC-anti-CD11b, FITC-anti-CD19, PE-anti-F4/80, PE-anti-Ly6G, PE-anti-CD8, or FITC-anti-CD4 antibodies) diluted in PBS for 1 h at room temperature in the dark. The cells were washed twice with PBS for flow cytometric analysis with a FACSC (BD Biosciences), and the data were analyzed with FlowJo (Tree Star Inc.)58 (link).
7
Annexin V-FITC/PI Apoptosis Assay
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Cell apoptosis was analyzed by Annexin V-FITC/Propidium Iodide (PI) Apoptosis Detection Kit according to the manufacturer's protocol (Sigma-Aldrich, Sigma Chemical Co., St. Louis, MO, USA). Briefly, H1299 cells were seeded in 6-well plates at a density of 1 × 104 cells/well and then treated with ZSD (4 mg/mL) for 12, 24, and 48 h, respectively, or different doses of ZSD. Then, cells were collected and centrifuged and resuspended in binding buffer. Afterwards, 5 μL Annexin V-FITC and 5 μL PI were added at room temperature for 15 min. Finally, apoptosis cells were analyzed by a flow cytometer (FACSC, BD Instruments Inc., USA) and FlowJo 7.1.0 software (Tree Star, Ashland, OR, USA).
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