Cell apoptosis was probed using the FITC Annexin V/7-AAD Apoptosis Detection Kit (BD Pharmingen, USA) according to the manufacturer's instructions. The ESCC cells were collected, washed with DPBS and Binding Buffer at 4°C, and resuspended in 200uL of Binding Buffer. A total of 1×106 cells were stained with 2.5uL of Annexin V-FITC and 5uL of 7-AAD. Cells were incubated with the staining solution at room temperature in the dark. Then cells were washed and acquired on a FACScan flow cytometer (BD FACSVerse™, USA).
Fitc annexin v 7 aad apoptosis detection kit
Manufactured by BD
Sourced in United States
The FITC-Annexin V/7-AAD apoptosis detection kit is a laboratory reagent used for the detection and quantification of apoptotic cells by flow cytometry. The kit contains Annexin V conjugated with the fluorescent dye FITC and the DNA-binding dye 7-AAD, which can identify cells in different stages of apoptosis.
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Lab products found in correlation
Cisplatin, by Promega (2 mentions)
5 5 6 6 tetrachloro 1 1 3 3 tetraethylbenzimidazolylcarbocyanine iodide jc 1, by Thermo Fisher Scientific (2 mentions)
Ldh dependent cytotoxic nonradioactive cytotoxicity assays, by Promega (2 mentions)
N acetylcysteine nac, by Merck Group (2 mentions)
Caspase 3 fluorometric assay kit, by Abcam (2 mentions)
Lc3b antibody, by Novus Biologicals (2 mentions)
Anti irf 1 antibody, by Santa Cruz Biotechnology (2 mentions)
β actin, by Merck Group (2 mentions)
Facscan flow cytometer, by BD (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
2 7 dichlorofluorescin diacetate dcfh da, by Merck Group (1 mentions)
Goat anti rabbit secondary antibody, by Thermo Fisher Scientific (1 mentions)
2 7 dichloro uorescin diacetate dcfh da, by Merck Group (1 mentions)
4 protocols using fitc annexin v 7 aad apoptosis detection kit
1
Evaluating Apoptosis in ESCC Cells
2
Apoptosis Detection by Flow Cytometry
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Apoptosis was detected by flow cytometric analysis using a FITC-Annexin V/7-AAD Apoptosis Detection Kit (BD, CA, USA). Briefly, cells (1 × 105 cells/well) were seeded in a 12-well plate. After culturing for 48 h, the treated cells were harvested, incubated with FITC-Annexin V and 7-AAD for 30 min at room temperature in the dark, and immediately analyzed by flow cytometry (FACSCalibur flow cytometer, BD, CA, USA). Each experiment was conducted at least three times.
3
Cytotoxicity and Apoptosis Assays
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Cisplatin, methyl thiazolyl tetrazolium (MTT), and LDH-dependent cytotoxic nonradioactive cytotoxicity assays were obtained from Promega (Madison, WI, USA). The LC3B antibody was purchased from Novus Biologicals (Littleton, CO, USA). N-acetylcysteine (NAC), β-actin, and 2,7-dichloro-fluorescin diacetate (DCFH-DA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). An anti-IRF-1 antibody was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The Caspase 3 Fluorometric Assay kit was obtained from BioVision (Milpitas, CA, USA). Goat anti-rabbit secondary antibodies were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). 5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraethyl-benzimidazolylcarbocyanine iodide (JC-1) was obtained from Invitrogen (Carlsbad, CA, USA). Assay kits for ATP, superoxide dismutase (SOD), and malondialdehyde (MDA) were obtained from Nanjing Jiancheng Bioengineering Institute. (Nanjing, China). The FITC-Annexin V/7-AAD apoptosis detection kit was obtained from BD Biosciences (USA).
4
Cytotoxicity Assays for Drug Screening
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Cisplatin, methyl thiazolyl tetrazolium (MTT), and LDH-dependent cytotoxic nonradioactive cytotoxicity assays were obtained from Promega (Madison, WI, USA). The LC3B antibody was purchased from Novus Biologicals (Littleton, CO, USA). N-acetylcysteine (NAC), β-actin, and 2,7-dichloro-uorescin diacetate (DCFH-DA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). An anti-IRF-1 antibody was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The Caspase-3 Fluorometric Assay kit was obtained from BioVision (Milpitas, CA, USA). Goat anti-rabbit secondary antibodies were purchased from Thermo Fisher Scienti c Inc. (Waltham, MA, USA). 5,5',6,6'-Tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide (JC-1) was obtained from Invitrogen (Carlsbad, CA, USA). Assay kits for ATP, superoxide dismutase (SOD), and malondialdehyde (MDA) were obtained from Nanjing Jiancheng Bioengineering Institute. (Nanjing, China). The FITC-Annexin V/7-AAD apoptosis detection kit was obtained from BD Biosciences (USA).
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