The assay was carried out using Annexin V FITC Kit (BD Pharmigen, Franklin Lakes, NJ, USA) according to manufacturer’s protocol to evaluate the apoptosis induction of rAF-IL12 towards cancer cells. Briefly, the seeded cells (2 x 105 cells/well) were infected for 72 h before the cell pellets were harvested and resuspended in the 1X Binding buffer prior to staining with FITC Annexin V and propidium iodide (PI) dyes as described by Najmuddin et al. [19 (link)]. The cell suspension was then allowed to stand in the dark at room temperature for 15 min before being analysed by NovoCyte Flow Cytometer (ACEA Biosciences Inc., San Diego, CA, USA) using NovoExpress® version 1.2.4 software (ACEA Biosciences Inc., San Diego, CA, USA).
Annexin 5 fitc kit
Manufactured by BD
Sourced in United States
The Annexin V-FITC kit is a laboratory reagent used for the detection and quantification of apoptosis, or programmed cell death, in cells. Annexin V is a calcium-dependent phospholipid-binding protein that has a high affinity for phosphatidylserine (PS), which is exposed on the outer membrane of cells undergoing apoptosis. The FITC (Fluorescein Isothiocyanate) label allows for the visualization and analysis of Annexin V-positive cells using flow cytometry or fluorescence microscopy.
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Lab products found in correlation
Facscalibur, by BD (14 mentions)
Facscan flow cytometer, by BD (11 mentions)
Facscalibur flow cytometer, by BD (8 mentions)
Cellquest software, by BD (5 mentions)
Accuri c6 flow cytometer, by BD (5 mentions)
Cytoflex, by Beckman Coulter (4 mentions)
Facscan, by BD (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Cell cycle analysis kit, by Beyotime (3 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (3 mentions)
Trypsin, by Thermo Fisher Scientific (3 mentions)
Annexin 5 fitc, by BD (3 mentions)
Accuri c6, by BD (3 mentions)
Novocyte flow cytometer, by Agilent Technologies (2 mentions)
Novoexpress version 1, by Agilent Technologies (2 mentions)
Flow cytometry, by Beckman Coulter (2 mentions)
Pierce ip lysis buffer, by Thermo Fisher Scientific (2 mentions)
Transwell plate, by Corning (2 mentions)
Facsaria, by BD (2 mentions)
Trypsin edta, by Nacalai Tesque (2 mentions)
220 protocols using annexin 5 fitc kit
1
Annexin V-FITC Apoptosis Assay
2
Apoptosis Induction in Cancer Cells
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The assay was carried out using Annexin V FITC Kit (BD Pharmigen, San Diego, CA, USA) according to manufacturer’s protocol to evaluate the apoptosis induction of rAF-IL12 towards cancer cells. The pellets of rAF-IL12-treated HT29 cells (72-h post-infection) were resuspended in 1x Binding Buffer prior to staining with FITC Annexin V and propidium iodide (PI) dyes. The cell suspension was then allowed to stand in the dark at room temperature for 15 mins before being analysed using the NovoCyte Flow Cytometer (ACEA Biosciences Inc., San Diego, CA, USA) and the NovoExpress® version 1.2.4 software (ACEA Biosciences Inc., San Diego, CA, USA) (Pawłowska et al., 2018 (link)).
3
Cell Cycle and Apoptosis Assay
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Cell cycle transition and cell apoptosis rates were investigated after cells were modified. The modified cells were treated with 0.5% trypsin and washed with PBS butter. For the cell cycle assay, cells were fixed in 1% paraformaldehyde and stained with 5 mg/ml propidium iodide. The apoptosis assay was performed using the BD Annexin V/FITC kit (BD, USA) and detected using a flow cytometer.
4
Cell Cycle and Apoptosis Analysis
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To analyse cell cycle distribution, CRC cells were fixed using 1% paraformaldehyde, washed with cold PBS, then, cells were incubated with 5 mg/ml propidium iodide (PI) and examined using flow cytometer. For detection of apoptosis, BD Annexin V/FITC kit (BD) was used to stain apoptotic CRC cells, which were detected using flow cytometer.
5
Cell Cycle and Apoptosis Profiling
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After washing with PBS, cells were fixed in 75% ethanol at 4°C overnight. Again washed with PBS, propidium iodide was used to stain cell in the dark at 37°C. Flow Cytometry (Beckman Coulter, Inc., Brea, CA) was used to measure the cell populations in different phases. To determine the cell apoptosis, Annexin V‐FITC Kit (Becton, Dickinson and Company, Franklin Lakes, NJ) was used according to the manufacturer's protocol. Finally, Flow Cytometry (Beckman Coulter, Inc.) was used to measure the cell populations in apoptosis. All assays were carried out biological independently in triplicate.
6
Quantifying Apoptosis in Rat Lungs
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Following paraffin embedding, six lungs of rats in each group was cut into 5 μm serial sections, and apoptosis was examined by TUNEL assay according to the manufacturer’s protocol (Roche Ltd., Shanghai, China). Briefly, Apoptotic indices (OD) were calculated as TUNEL-positive cells by Image-Pro plus 6.0 software (Media Cybernetics, Inc. Rockville, MD).
The apoptosis of thymocytes was detected using flow cytometry. About 1×106 thymocytes per milliliter were harvested and resuspended in 100 μL binding buffer. Annexin V coupled with FITC was added according to the AnnexinV-FITC kit (Becton Dickinson, USA). Following 30 mins of incubation at room temperature in darkness, 50 μg/mL Propidium Iodide was added and incubated for 1 min before FACS analysis (lymC6 flow cytometer, Becton Dickinson, USA). Data from 1×104 cells were collected and analyzed in each sample.
The apoptosis of thymocytes was detected using flow cytometry. About 1×106 thymocytes per milliliter were harvested and resuspended in 100 μL binding buffer. Annexin V coupled with FITC was added according to the AnnexinV-FITC kit (Becton Dickinson, USA). Following 30 mins of incubation at room temperature in darkness, 50 μg/mL Propidium Iodide was added and incubated for 1 min before FACS analysis (lymC6 flow cytometer, Becton Dickinson, USA). Data from 1×104 cells were collected and analyzed in each sample.
7
Annexin V-FITC Apoptosis Assay
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Apoptosis was evaluated by cytofluorometry using the Annexin V-FITC Kit (Becton Dickinson, Mountain View, CA, USA). Briefly, following the indicated treatments, cells were centrifuged at 100 × g for 5 min, washed twice with cold PBS, and then resuspended in 500 µL binding buffer. The cells were then labeled with Annexin V-FITC (5 µL) and propidium iodide (PI, 5 µL) for 15 min in the dark at room temperature and analyzed using a FACS Calibur flow cytometer (Becton Dickinson, Mountain View, CA, USA).
8
Apoptosis Assessment of hRECs Using Annexin V/FITC
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The Annexin V/FITC kit [Becton, Dickinson, and Co., (BD) Biosciences, San Jose, CA, USA] was employed to label apoptotic cells in accordance with the manufacturer’s instructions, and the cells were monitored by FCM. The hRECs were digested with trypsin without ethylenediamine tetraacetic acid (EDTA), and then were collected in medium and precipitated by a 5 min centrifugation at 500 ×g. After being washed and resuspended with 1× binding buffer, hRECs were mixed with 5 µL annexin V-FITC, and further incubated in the dark at room temperature for 10 min. The hRECs were stained with 10 µL of propidium iodide (PI; 20 µg/mL) for 5 min in the dark, and finally applied for florescent detection. Each experiment was repeated 3 times.
9
Annexin V Apoptosis Assay Protocol
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Apoptosis was assessed by the AnnexinV-FITC Kit (Becton Dickinson, San Diego, CA, USA). The cells were incubated with Uncaria tomentosa extract for 72 h in 12-well plates. After the exposure, the culture medium was removed and the cells were harvested by trypsinization with 0.05% Trypsin-EDTA, and centrifuged at 2000 rpm for 10 min. The analysis by flow cytometery was performed within 1 h by counting 5000 gated events per each sample. The data were further analyzed using CELL-Quest software on FACS Calibur flow cytometer and the Cell Quest software (BD Biosciences, San Jose, CA, USA). The results were presented as rates of early apoptotic (Annexin V-positive) and late apoptotic (Annexin V and PI-positive) cells.
10
Annexin V Apoptosis Analysis
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Apoptosis analysis was evaluated by cytofluorometry using the annexin V‐FITC kit (Becton Dickinson) according to the manufacturer’s instructions. Apoptosis was determined by FACSCalibur flow cytometry (Becton Dickinson).
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