Cell death via apoptosis was determined using Annexin V‐APC/7‐AAD Apoptosis Kit (BD Biosciences, San Diego, CA) and quantified by flow cytometry as described previously 14 , 15 . Briefly, at the end of the treatment time, DU145 cells were washed once with PBS and harvested in a 0.5% trypsin/EDTA solution at 37°C, centrifuged at 220g for 5 min and then immediately resuspended in the physiological buffer provided in the kit. Cells (1 × 105 cells/500 μL) were then maintained in the dark for 15 min at room temperature with both Annexin V, allophycocyanin conjugate (APC) and 7‐amino‐actinomycin D (7‐AAD) accordingly, after which the samples were analyzed immediately by BD Biosciences FACSCalibur flow cytometer (San Jose, CA). The results were quantified using the CellQuest software (BD Biosciences, San Jose, CA). Q2 plus Q4 area were calculated as apoptosis ratio.
Annexin 5 apc 7 aad apoptosis kit
Manufactured by BD
Sourced in United States
The Annexin V-APC/7-AAD apoptosis kit is a laboratory reagent used for the detection and quantification of apoptosis in cell samples. It contains Annexin V conjugated to the fluorescent dye APC (Allophycocyanin) and the DNA-binding dye 7-AAD (7-Aminoactinomycin D). This kit enables the simultaneous identification of early and late apoptotic cells through flow cytometric analysis.
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Lab products found in correlation
Accuri c6, by BD (2 mentions)
Cycletest plus dna reagent kit, by BD (2 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Cellquest software, by BD (1 mentions)
Annexin 5 fitc pi apoptosis kit, by MultiSciences Biotech (1 mentions)
Cytexpert v2, by Beckman Coulter (1 mentions)
Cytoflex s, by Beckman Coulter (1 mentions)
Cell cycle kit, by Keygen Biotech (1 mentions)
8 protocols using annexin 5 apc 7 aad apoptosis kit
1
Annexin V Apoptosis Assay for DU145 Cells
2
Assessing Cell Apoptosis via Flow Cytometry
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Cell apoptosis was assessed by using Annexin V APC/7-AAD apoptosis kit (BD Pharmingen, USA). Samples were trypsinized, washed with ice-cold PBS, and stained with 5 μL Annexin V APC and 5 μL 7-AAD according to the manufacturer’s instructions. Analysis was carried out immediately on a FC500 flow cytometry system and FlowJo software. All samples were assayed in triplicate. The predicted results were that regular cells (APC−/7-AAD−), necrotic cells (APC+/7-AAD+), and apoptotic cells (APC+/7-AAD−).
3
Quantifying Cell Apoptosis via Flow Cytometry
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Cell apoptosis was determined using Annexin V-FITC/PI apoptosis kit (Multi Sciences) or Annexin V-APC/7-AAD apoptosis kit (BD Pharmingen, San Diego, CA, USA). According to the manufacturer’s instructions, cells (5 × 105) and incubated with Annexin V and PI or 7-AAD in the dark for 15 min. Apoptotic cells were detected with a flow cytometer (Beckman Coulter Inc, Miami, FL, USA). Three independent experiments were performed.
4
Annexin V-APC/7-AAD Apoptosis Assay
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Annexin V-APC/7-AAD apoptosis kit Flow was purchased from BD Bioscience (USA, Catalogue No. 550474). Washed cells twice with cold PBS and then resuspended cells in 1X Binding Buffer at a concentration of 1 × 106 cells/ml. Transfered 100 μl of the solution (1 × 105 cells) to a 5 ml culture tube. Added 5 μl of APC Annexin V (for one and two color analysis) and 5 μl of 7-AAD (for two color analysis only). Gently vortexed the cells and incubated for 15 min at RT (25 °C) in the dark. Added 400 μl of 1X Binding Buffer to each tube. Analyzed by flow cytometry within 1 h.
5
Annexin V-APC/7-AAD Apoptosis Assay
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The treated cells were collected and transferred into a centrifuge tube. After centrifugation (1,000 × g, 4°C, 5 min), the cells were resuspended in 100 µl binding buffer. An Annexin V-APC/7-AAD apoptosis kit (BD Biosciences) was used for double staining in the dark. The apoptosis rate of the HEI-OC1 cells was measured by flow cytometry (CytoFLEX S; Beckman Coulter, Inc.). CytExpert (v2.0; Beckman Coulter, Inc.) was used to analyze the data.
6
Cell Cycle and Apoptosis Analysis
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The cell cycle kit (KeyGEN, Nanjing, China) and Annexin V-APC/7-AAD apoptosis kit (BD Pharmingen, New Jersey, USA) were utilized to detect the cell cycle distribution and apoptosis rates, respectively. The procedure was performed according to a previous description [20 (link)].
7
Cell Cycle and Apoptosis Analysis
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For the cell cycle analysis, the transfected cells were stained with propidium iodide by the Cycletest Plus DNA reagent kit (BD Biosciences, USA) and then measured by flow cytometry (BD Biosciences Accuri C6, USA). The ratios of cells in G0/G1, S, and G2 phases were counted and compared. To detect cell apoptosis, cells were stained using an annexin V-APC/7AAD apoptosis kit (BD Biosciences, USA) and analyzed using flow cytometry. The ratio of early apoptotic cells to late apoptotic cells was compared to the values obtained for the controls in each experiment.
8
Cell Cycle and Apoptosis Analysis
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For the cell cycle analysis, the transfected cells were stained with propidium iodide by the Cycletest Plus DNA reagent kit (BD Biosciences, USA) and then measured by flow cytometry (BD Biosciences Accuri C6, USA). The ratios of cells in G0/G1, S, and G2 phases were counted and compared. To detect cell apoptosis, cells were stained using an annexin V-APC/7AAD apoptosis kit (BD Biosciences, USA) and analyzed using flow cytometry. The ratio of early apoptotic cells to late apoptotic cells was compared to the values obtained for the controls in each experiment.
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