Apoptosis assay was performed according to the manual of BD Annexin V-APC apoptosis detection Kit (Cat. 550474, BD Biosciences, Franklin Lakes, NJ USA). CAFs were detached, washed twice with PBS and resuspended in 1×Annexin binding Buffer. These CAFs were stained with annexin V conjugated with APC at room temperature in the dark for 15–20 min. After incubation, cells were washed twice with 1 × Annexin binding Buffer and resuspended in 1 × Annexin binding Buffer containing Propidium Iodide and stained cells(apoptotic cells) were detected by flow cytometry immediately.
Apc annexin 5 apoptosis detection kit
Manufactured by BD
Sourced in United States
The APC Annexin V Apoptosis Detection Kit is a lab equipment product designed to detect and quantify apoptosis in cell populations. It utilizes Annexin V, a protein that binds to phosphatidylserine, a component exposed on the cell surface during apoptosis. The kit provides a fluorescent APC-conjugated Annexin V reagent to label apoptotic cells, which can then be detected and analyzed using flow cytometry or fluorescence microscopy.
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Lab products found in correlation
Facscanto 2, by BD (5 mentions)
Facscalibur flow cytometer, by BD (5 mentions)
Lsrfortessa, by BD (4 mentions)
Facscalibur, by BD (3 mentions)
Facsverse flow cytometer, by BD (3 mentions)
Facsuite software, by BD (3 mentions)
Flow cytometry, by BD (2 mentions)
Facscan, by BD (2 mentions)
Lsr 2 flow cytometer, by BD (2 mentions)
Facsaria, by BD (2 mentions)
Facsaria flow cytometer, by BD (2 mentions)
Cell cycle analysis kit, by BD (1 mentions)
Fasc calibur mt flow cytometer, by BD (1 mentions)
Fcs express 4, by De Novo Software (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Propidium iodide, by BioLegend (1 mentions)
Rnase a, by Sangon (1 mentions)
Canto 2, by BD (1 mentions)
Facs flow cytometer, by BD (1 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
58 protocols using apc annexin 5 apoptosis detection kit
1
Annexin V-APC Apoptosis Assay for CAFs
2
Apoptosis Induction and Detection
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Cells were induced apoptosis by treatment of 1.0 μM Doxorubicin. Apoptosis assay was performed according to the manual of BD Annexin V-APC apoptosis detection Kit (Cat. 550474, BD Biosciences, Franklin Lakes, NJ, USA). Briefly, cells were washed twice with PBS and then resuspended in 1× Annexin binding Buffer. These cells were stained with Annexin V conjugated with APC at room temperature in the dark for 20 min. Stained cells were detected by flow cytometry.
3
Annexin V-APC Apoptosis Assay in H9c2 Cells
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The level of apoptosis in the treated H9c2 cells was determined using an Annexin V-APC Apoptosis Detection kit (BD Biosciences) according to the manufacturer's instructions. The data were analyzed using a flow cytometer and Image-Pro Plus 6.0 used to analyze.
4
Quantifying Apoptosis in Cancer Cells
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The extent of apoptosis was measured with an Annexin V-APC Apoptosis Detection kit (BD Biosciences) according to the manufacturer′s instructions. Untransfected and transfected GBC-SD, SGC-996, and NOZ cells were collected, washed with cold PBS twice, gently resuspended in 100 μL of 1× binding buffer containing 2.5 μL APC-conjugated annexin-V and 1 μL of 100 μg/mL PI, and then incubated at room temperature in the dark for 15 min. The stained cells were analyzed by flow cytometry (BD Biosciences). The experiments were performed in triplicate.
5
Quantifying p75NTR+ Cells and Cell Cycle Dynamics in ESCC
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Flow cytometry was performed to detect the ratio of p75NTR+ cells in ESCC cells. Briefly, Cells were trypsinized and collected by centrifugation. Then cells were washed and suspended in buffer and incubated with p75NTR-PE antibody (BD Bioscience) at 4°C for 2h. The ratio of p75NTR+ cells in each samples were analyzed by FASC Calibur MT flow cytometer (BD Bioscience).
Cell cycle was detected by flow cytometry using a cell cycle analysis kit (BD Bioscience). In brief, at least 1*106 cells were harvested and washed, then cells were fixed in ice-cold 70% ethanol for at least 2h at 4°Cwashed the cells again and stained with a solution containing 50μg/ml PI and 50 RNase at room temperature for 30 min. Cell cycle was analyzed with a FACS Calibur MT flow cytometer (BD Bioscience).
Cell apoptosis was detected by flow cytometry using an Annexin V-APC apoptosis detection kit (BD Bioscience). In brief, cells were harvested and washed, then cells were suspended in binding buffer and incubated with Annexin V-APC and propidium iodide (PI) at 4°C, the cells were double stained with Annexin V-APC and PI according to the manufacturer's instructions. Early apoptosis and the late apoptosis were determined by Annexin V+/PI- staining and Annexin V+/PI+ staining, respectively. The percentage of apoptosis cells in each sample was examined using the FASC Calibur MT flow cytometer (BD Bioscience).
Cell cycle was detected by flow cytometry using a cell cycle analysis kit (BD Bioscience). In brief, at least 1*106 cells were harvested and washed, then cells were fixed in ice-cold 70% ethanol for at least 2h at 4°Cwashed the cells again and stained with a solution containing 50μg/ml PI and 50 RNase at room temperature for 30 min. Cell cycle was analyzed with a FACS Calibur MT flow cytometer (BD Bioscience).
Cell apoptosis was detected by flow cytometry using an Annexin V-APC apoptosis detection kit (BD Bioscience). In brief, cells were harvested and washed, then cells were suspended in binding buffer and incubated with Annexin V-APC and propidium iodide (PI) at 4°C, the cells were double stained with Annexin V-APC and PI according to the manufacturer's instructions. Early apoptosis and the late apoptosis were determined by Annexin V+/PI- staining and Annexin V+/PI+ staining, respectively. The percentage of apoptosis cells in each sample was examined using the FASC Calibur MT flow cytometer (BD Bioscience).
6
Cell Cycle and Apoptosis Analysis
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For cell cycle detection, infected cells cultured in DMEM from all groups were digested with 0.25% trypsin (Invitrogen; Thermo Fisher Scientific, Inc.), collected by centrifuging at 377 × g for 6 min at 4°C and washed once with phosphate-buffered saline (PBS; Shanghai Sangon Biotech Co.). Cells were fixed with ice-cold 75% ethanol at 4°C overnight. Subsequently, cells were centrifuged (377 × g for 6 min at 4°C) and ethanol was removed by washing with PBS three times. Cells were slightly resuspended with 300 µl PBS and treated with 50 µg/ml RNase A (Shanghai Sangon Biotech Co., Ltd.) for 30 min at 37°C. Cells were stained with propidium iodide (PI; BioLegend, Inc., San Diego, CA, USA) in the dark for 15 min at 4°C and detected using a flow cytometer (BD Biosciences, San Jose, CA, USA). Data were analyzed by FCS Express 4 (De Novo Software, Los Angeles, CA, USA).
Annexin V-APC Apoptosis Detection kit (BD Biosciences) was used to detect cell apoptosis. Infected cells were digested with 0.25% trypsin-EDTA (Invitrogen; Thermo Fisher Scientific, Inc.) and collected by centrifuging at 377 × g for 6 min at 4°C, then washed once with PBS. Cells were added to APC-Annexin V and PI in the dark for 15 min at 25°C after being slightly resuspended with 1X Binding Buffer. A total of 400 µl 1X Binding Buffer was added and the cells were detected using a flow cytometer.
Annexin V-APC Apoptosis Detection kit (BD Biosciences) was used to detect cell apoptosis. Infected cells were digested with 0.25% trypsin-EDTA (Invitrogen; Thermo Fisher Scientific, Inc.) and collected by centrifuging at 377 × g for 6 min at 4°C, then washed once with PBS. Cells were added to APC-Annexin V and PI in the dark for 15 min at 25°C after being slightly resuspended with 1X Binding Buffer. A total of 400 µl 1X Binding Buffer was added and the cells were detected using a flow cytometer.
7
Multiparametric Flow Cytometry Analysis
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A BD LSRII Flow Cytometer was used in all analyses, and FlowJo v10 was used for data analysis. For measurements of apoptosis, 0.5×106 cells were stained with an Annexin V-APC Apoptosis Detection Kit (BD Biosciences, San Diego, USA) based on provided directions.
For cell cycle analyses, 0.5×106 cells were fixed overnight at 4°C in 75% ethanol, washed thrice in PBS, and stained using propidium iodide for 20 minutes.
For immunophenotyping analyses, BM, PB, and spleen cells (0.5×106) were collected, washed thrice using PBS, and stained at 4°C with antibodies specific for CD11b, CD117, CD45.1, CD45.2, and Gr-1 (BD Biosciences) for 20 minutes. After two additional washed, cells were then fixed in a fixation buffer prior to analysis.
For cell cycle analyses, 0.5×106 cells were fixed overnight at 4°C in 75% ethanol, washed thrice in PBS, and stained using propidium iodide for 20 minutes.
For immunophenotyping analyses, BM, PB, and spleen cells (0.5×106) were collected, washed thrice using PBS, and stained at 4°C with antibodies specific for CD11b, CD117, CD45.1, CD45.2, and Gr-1 (BD Biosciences) for 20 minutes. After two additional washed, cells were then fixed in a fixation buffer prior to analysis.
8
Quantifying Apoptosis by Flow Cytometry
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Apoptosis was measured by staining with annexin V–APC and Propidium Iodide (PI)-phycoerythrin (PE) (Annexin V-APC Apoptosis Detection kit, BD Pharmingen) followed by flow cytometry on a FACS flow cytometer (BD, Canto II). All experiments were performed in triplicate, and results were calculated as the mean ± S.D.
9
Quantifying Glioblastoma Apoptosis in IFN-DCs
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Glioblastoma cells stained with 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE) fluorescent dye (2 μM; Molecular probes, Inc., Eugene, OR, USA) were incubated with IFN-DCs at a ratio DCs:tumor cells 10:1 for 18 h. Percentage of tumor cells apoptosis was measured using annexin V-APC apoptosis detection kit (BD Pharmingen™, USA). Briefly, cells were washed with PBS and labeled with APC-conjugated annexin V and propidium iodide (PI) for 15 min at room temperature, followed by analysis on a FACSCalibur flow cytometer. A minimum of 10,000 events within the CFSE-positive gate region were collected for each sample.
10
Annexin V-based Apoptosis Assay for PDAC
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Apoptosis assay was performed by AnnexinV/APC apoptosis detection kit (BD biosciences, San Jose, CA, USA). 0.1–0.2 × 106 PDAC cells were treated with increasing concentrations of PVT (0–7.5 μM) for 24 h. At the desired time points, cells were trypsinized and then analyzed for Annexin and PI staining using flow cytometry (BD LSR Fortessa, BD biosciences, San Jose, CA, USA). The flow cytometry data were integrated into the FlowJo software and % apoptotic cells were quantified. All the experiments were repeated independently for 2–3 times.
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