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Annexin 5 fitc pi

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Annexin V-FITC/PI is a fluorescent staining kit used to detect and quantify apoptosis in cells. It contains Annexin V conjugated with fluorescein isothiocyanate (FITC) and propidium iodide (PI). Annexin V binds to phosphatidylserine, which is exposed on the cell surface during apoptosis, while PI binds to DNA in cells with compromised cell membranes, indicating late apoptosis or necrosis.

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49 protocols using annexin 5 fitc pi

1

Evaluating Cell Proliferation and Apoptosis

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To assess cell proliferation, flow cytometric analysis was performed using PE-conjugated monoclonal anti-Ki-67 antibody (clone: 20Raj1, Bioscience) and apoptosis performed by Caspase 3 kit (BD Biosciences, San Diego, CA). For intracellular staining, cells were collected and washed twice with ice-cold PBS, resuspended with BD cytofix/cytoperm solution and incubated at 4°C for 20 minutes, cells were washed twice using Perm/Wash buffer and incubated with anti-Ki-67 or anti-caspase 3 antibody at room temperature for 20 minutes and analyzed by BD FACS Calibur with FlowJo (7.6.1) software for acquisition and analysis.
Also, flow cytometric analysis was performed using Annexin V-FITC/PI staining. Following 48h treatment, cells were collected and rinsed twice with ice-cold PBS. Then cells washed and incubated with Annexin V-FITC/PI (eBiosciences) for 15 min at room temperature. Cells were analyzed by BD FACS Calibur with FlowJo (7.6.1) software for acquisition and analysis.
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2

Annexin V-FITC/PI Flow Cytometry

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DHL cells were treated with designated drugs for indicated times and then harvested and subjected to flow cytometric analysis using Novocyte (ACEA Bioscience, San Diego, CA, USA) after Annexin V-FITC/PI (eBioscience, San Diego, California, USA) staining following the manufacturer’s instruction. All experiments were performed in triplicate, and the data were indicated as mean ± SD.
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3

Annexin-V-FITC/PI Apoptosis Assay

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To assess the apoptosis, the cells were treated with chidamide or apatinib alone or in combination for 48 h and stained with the Annexin-V-FITC/PI (eBioscience, San Diego, CA, USA) for 15 min at room temperature in the dark according to the manufacturer’s instructions. The cells were then analyzed by NovoCyte flow cytometer and the data were analyzed using the NovoExpress software (ACEA Biosciences, Inc., San Diego, CA, USA) to determine the percentage of the Annexin-V positive (apoptotic) cells.
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4

Annexin V-FITC/PI Apoptosis Assay

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An apoptosis detection kit, annexin V‐FITC/PI obtained from eBioscience, was employed to assess cell apoptosis. Following staining protocols as per the provided instructions, the cells were examined via flow cytometry.
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5

Apoptosis Analysis in AML Cell Lines

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To assess apoptosis, OCL-AML2, OCL-AML3, MOLM-13, and MV4-11 cells were cultured and treated with different doses of ABT-199 or HHT alone or in combination for 24 h or 48 h and then double-labeled with Annexin-V-FITC/PI (eBioscience, San Diego, California, USA) for 20 min at room temperature in the dark according to the manufacturer’s instructions. The stained cells were analyzed with a NovoCyte flow cytometer (ACEA Biosciences, Inc.) with NovoExpress software. Apoptotic cells were defined as Annexin-V positive.
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6

Apoptosis Quantification by Flow Cytometry

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Apoptosis was measured using Annexin V-FITC/PI (Ebioscience, San Diego, USA) dual staining by flow cytometry. Briefly, cells (1 × 105 /well) were seeded into 6-well plates and infected with lentivirus mediated FAK shRNA-1 or sh-ctrl with or without exposed to 5-FU for 24 h. Cells were harvested and washed in cold FACS buffer (PBS containing 2% FBS), and labeled with Annexin V-FITC for 30 min at 4 °C in the dark and then with PI. The stained cells were analyzed by flow cytometry (LSRFortessa, Becton Dickinson, San Jose, CA, USA). The filters used in the flow cytometry were: DAPI: 450 BP 40, FITC: 525 BP40.
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7

Apoptosis Assessment in Leukemia Cells

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To assess apoptosis, CD34+CD38 KG1α or Kasumi cells were cultured as described above for 24, 48, or 72 h with or without chidamide, Ara-C, DNR, or IDA, then double labeled with Annexin V-APC/PI (eBioscience, San Diego, CA, USA) for 15 min at room temperature in the dark according to the manufacturer’s instructions. Primary samples were stained with hCD34-APC (eBioscience, USA) and Annexin V-FITC/PI to assess the apoptosis of CD34+ primary cells induced by chidamide or IDA alone or the two drugs in combination. The stained cells were analyzed by flow cytometry (FACS Fortessa, BD Biosciences). Apoptotic cells were defined as Annexin V positive.
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8

Apoptosis Assay of miR-34a-5p Transfected Cells

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Forty-eight hours after transient transfection of KatoIII cells with miR-34a-5p mimic or negative control, the cells were collected and stained with Annexin V‑FITC/PI according to the manufacturer’s instructions (eBioscience; Cat. no. BMS500FI/100) to assay apoptosis. The apoptosis level of stably transfected cells was assessed using Annexin V‑PE/7-AAD (BioLegend; Cat. no. 640,908) in addition to Annexin V‑FITC/PI staining. Flow cytometry was performed to detect the apoptosis level of the transfected cells. The data was analyzed by FlowJo software 7.6.1 (Tree star, Inc., San Carlos, CA, USA).
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9

Apoptosis Quantification in HA-SMCs

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In vitro assessment of apoptosis in HA-SMCs, ImageStreamχ MK II imaging cytometer (Amnis Corp., Seattle, WA, USA) via annexin V-FITC/PI (eBioscience, BMS500FI, San Diego, USA) staining was used to observe the induction of apoptosis. Cells in the lower and upper right quadrant indicated early apoptosis (annexin-positive/PI-negative) and late apoptosis (annexin-positive/PI-positive), respectively. The percentage of apoptosis = (early apoptosis + late apoptosis)/total cells.
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10

Apoptosis Analysis of HL-60 Cells

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Flow cytometric analysis of HL-60 cells double-stained with apoptosis annexin V-FITC/PI (eBioscience, Vienna, Austria) was used to determine cell apoptosis. Briefly, HL-60 cells were seeded in 24-well plates and incubated with e-As4S4/1:15–60 at different concentrations of As4S4 for 6–48 h. After centrifugation and washing with PBS, cells were resuspended in binding buffer. Then, annexin V-FITC and PI were added, followed by a 10-min incubation at room temperature, and were analyzed using a flow cytometer (Accuri C6; BD Company, Franklin Lakes, NJ). In total, 1.5 × 104 cells were measured and the data acquired were analyzed with the CFlow Plus software.
Apoptotic cells were also examined by microscopy following Hoechst 33342 (Beyotime Biotechnology, Haimen, China) staining. Cells were seeded in 24-well plates and a suspension of e-As4S4/1:15–60 at 40 mg/L of As4S4 was added. After 48 h incubation, the cells were harvested in a 1.5-mL concentration tube, washed twice with PBS, and suspended in 1 mL medium. Hoechst 33342 at 5 μL/mL was added to the tube and incubated for 25 min at 4 °C in the dark. Then the cells were washed twice with PBS to remove unbound dye and imaged with a fluorescence microscope (IX71, Olympus, Tokyo, Japan).
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