The Annexin V/FITC and propidium iodide (PI) apoptosis detection kit (Becton-Dickinson, Franklin Lakes, NJ, USA) was used to quantitatively measure the phosphatidylserine in apoptotic cells. Briefly, transfected cells (siCtl, siPCDH1) (5 × 105 per well) were seeded into 6-well plates. After 24 h, the cells were harvested and washed three times with ice-cold phosphate-buffered saline (PBS) (pH 7.2). After washing, each sample was centrifuged at 1300 rpm for 3 min at 4 °C. Annexin V/FITC and PI double-staining were performed according to manufacturer instructions. Apoptosis was analyzed on a FACScan flow cytometer (Becton-Dickinson, Heidelberg, Germany) and Annexin V-positive, PI-negative cells were scored as apoptotic (Fig 4 ). Double-stained cells were considered to be necrotic or late apoptotic.
Annexin 5 fitc and propidium iodide apoptosis detection kit
Manufactured by BD
Sourced in United States
The Annexin V-FITC and propidium iodide (PI) apoptosis detection kit is a laboratory product used to detect and quantify apoptosis, a form of programmed cell death. The kit contains Annexin V, a protein that binds to phosphatidylserine, and propidium iodide, a fluorescent dye that binds to DNA. These components are used to identify and differentiate between viable, early apoptotic, and late apoptotic/necrotic cells.
Automatically generated - may contain errors
Lab products found in correlation
Facscan, by BD (1 mentions)
Cytomics fc500 flow cytometer, by Beckman Coulter (1 mentions)
Facsverse, by BD (1 mentions)
Facsverse flow cytometer, by BD (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Facscanto system, by BD (1 mentions)
Flow cytometry instrument, by BD (1 mentions)
Cellquest pro software version 5, by BD (1 mentions)
Propidium iodide staining solution, by BD (1 mentions)
Annexin 5 fluorescein isothiocyanate fitc, by BD (1 mentions)
Accuri c6, by BD (1 mentions)
12 protocols using annexin 5 fitc and propidium iodide apoptosis detection kit
1
Quantitative Apoptosis Measurement Using Annexin V/PI
2
Evaluating HUVEC Apoptosis via Annexin V/PI
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The apoptosis of HUVECs was evaluated by using Annexin V/FITC and propidium iodide (PI) apoptosis detection kit (Becton Dickinson, Pasadena, USA) according to the manufacturer’s instructions. Briefly, treated cells were collected, and were suspended in Annexin-binding buffer, followed by staining with Annexin V- FITC/PI for 15 min in the dark at room temperature. Subsequently, the stained cells were analyzed by flow cytometry using the CYTOMICS FC 500 flow cytometer (Beckman Coulter, Brea, USA).
3
Cell Proliferation and Apoptosis Assay
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The CCK-8 test was used to determine cell proliferation. 1 × 103 cells were seeded into 96-well plates and cultivated for 24, 48, and 72 hours. The CCK-8 solution was then added to a 96-well plate and incubated for additional two hours. At 450 nm, the absorbance was determined. Flow cytometry was used to detect apoptosis using an Annexin V-FITC and propidium iodide (PI) apoptosis detection kit (Becton Dickinson). We collected 1 × 106 cells and suspended them in Annexin-binding buffer. Following that, cells were incubated for 15 minutes at room temperature in the dark with Annexin V-FITC and PI and immediately analyzed using the flow cytometry software equipped in the machine (BD FACSVerse; Becton-Dickinson and Company).
4
Annexin V-FITC and PI Apoptosis Assay
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Cell apoptosis was determined by an Annexin V/FITC and propidium iodide (PI)
apoptosis detection kit (Becton Dickinson, USA) according to the manufacturer's
instructions. Briefly, 48 h after treatment and transfection, cells were collected
and suspended in Annexin-binding buffer. The cells were then incubated with Annexin
V-FITC and PI for 15 min in the dark at room temperature. Flow cytometry was
performed to analyze the apoptotic percentage of cells.
apoptosis detection kit (Becton Dickinson, USA) according to the manufacturer's
instructions. Briefly, 48 h after treatment and transfection, cells were collected
and suspended in Annexin-binding buffer. The cells were then incubated with Annexin
V-FITC and PI for 15 min in the dark at room temperature. Flow cytometry was
performed to analyze the apoptotic percentage of cells.
5
Apoptosis Evaluation in 3D Lung Cancer Spheroids
6
Annexin V-FITC Apoptosis Detection
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Apoptotic cells were identified by using the Annexin V-FITC and propidium iodide (PI) apoptosis detection kit (BD, San Diego, USA) according to the manufacturer’s instructions. After being washed twice with PBS, the cells were resuspended in 400 µL 1 × binding buffer and stained with 5 µL Annexin V-FITC and 10 µL PI for 15 min at 4°C in the dark. Apoptosis was analyzed with a BD FACS CantoTM II flow cytometer (BD Bioscience, California, USA).
7
Evaluating CD8+ T Cell Apoptosis
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CD8+ T cells were isolated from splenocytes of the Anchored-GM-CSF vaccine group (on day 28 after tumor cell injection) using immunomagnetic microbeads. CD8+ T cells were cultured with MB49 cells on 24-well plates and treated with anti-PD-1 or IgG antibody for 72 hours. Then, cell apoptosis was measured by flow cytometry using an annexin V/FITC and propidium iodide (PI) apoptosis detection kit (BD Bioscience). Annexin V(+)/PI(-) and annexin V(+)/PI(+) indicated apoptosis and necrosis, respectively.
8
Apoptosis Quantification by Flow Cytometry
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Flow cytometry analysis was performed using a double staining Annexin V-FITC and propidium iodide (PI) apoptosis detection kit (BD Biosciences, USA), according to the manufacturer’s protocol for in vitro analysis. After processing using the kit, apoptosis analysis was carried out using a FACS CantoTM system (BD Biosciences, USA) at the FACS Core Facility, Tzu-chi Hospital Research Center, Taiwan. The apoptotic cells were gated (n=10,000 cells), and the proportion of apoptotic cells was calculated by adding the numbers of cells in the Q2 (late apoptosis) and Q4 (early apoptosis) quadrants.
9
Quantifying Cell Apoptosis by Flow Cytometry
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Apoptotic cells were detected by staining with Annexin-V-FITC and propidium iodide (PI) Apoptosis Detection Kit (BD, USA). The cell cycle stage measured by flow cytometry instrument (Becton-Dickinson, USA). The Annexin V-FITC−/PI− cells were considered to normal healthy cells. Cells stained with Annexin V-FITC+/PI− were regarded as early apoptosis cells. The Annexin V-FITC+/PI+ cells were regarded as a measure of late apoptosis. The Annexin V-FITC−/PI+ cells were considered as necrotic cells. Both early (Annexin V-FITC+/PI−) and late (Annexin V-FITC+/PI+) apoptotic cells were included in the cell death analyses.
10
Evaluation of Treg Apoptosis
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Tregs were cultured for 2, 4 or 6 days as previously described, and then washed and resuspended at a concentration of 1×106 cells/ml. To evaluate the apoptosis of Tregs, cells were detected using flow cytometry by an Annexin-V/FITC and propidium iodide (PI) apoptosis detection kit (BD Bioscience). AnnexinV(+)/PI(-) and AnnexinV(+)/PI(+) were defined as early apoptosis and late apoptosis, respectively.
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