The cells from each group were placed into 6-well plates at 3 × 105 cell/well and cultured overnight. Annexin V-FITC Apoptosis Detection Kit purchased from Sungene Biotech (Tianjin, China) was employed for annexin V-FITC/propidium iodide (PI) double-staining, following the manufacturer’s protocol. Apoptotic cells were detected on a BD AriaIII flow cytometry system (BD Biosciences, Franklin Lake, NJ, USA).
Annexin 5 fitc apoptosis detection kit
Manufactured by Sungene Biotech
Sourced in China
The Annexin V-FITC Apoptosis Detection Kit is a laboratory tool used to detect and quantify apoptosis, a type of programmed cell death. The kit contains Annexin V, a protein that binds to a cell membrane component exposed during apoptosis, conjugated with the fluorescent dye FITC. This allows the identification and measurement of apoptotic cells using flow cytometry or fluorescence microscopy.
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Lab products found in correlation
Trypsin edta solution, by Beyotime (1 mentions)
Mitochondrial membrane potential assay kit with jc 1, by Beyotime (1 mentions)
Anti bax, by Bioss Antibodies (1 mentions)
Anti bcl 2, by Bioss Antibodies (1 mentions)
Anti caspase 3, by Bioss Antibodies (1 mentions)
β actin antibody, by Bioss Antibodies (1 mentions)
Dulbecco modified eagle medium (dmem), by Corning (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Beyotime (1 mentions)
Dimethyl sulfoxide (dmso), by Beyotime (1 mentions)
Penicillin streptomycin solution, by Beyotime (1 mentions)
Accuri c6, by BD (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Facsaria 3, by BD (1 mentions)
Cytoflex flow cytometer, by Beckman Coulter (1 mentions)
Facscanto, by BD (1 mentions)
β actin, by Wuhan Sanying Biotechnology (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Bca protein assay kit, by Leagene (1 mentions)
14 protocols using annexin 5 fitc apoptosis detection kit
1
Annexin V-FITC Apoptosis Assay
2
Tet-induced Apoptosis and Cell Cycle Arrest in HT-29 Cells
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Tet (purity 99.1%) was purchased from Alphabio Biotechnology Co. Ltd (Tianjin, China). The HT-29 cell line was obtained from Cell Bank of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (Shanghai, China). DMEM was purchased from Corning Cellgro Inc. (Herndon, VA, U.S.A.) and the fetal bovine serum (FBS) was obtained from Biological Industries Technologies (Kibbutz Beit Haemek, Israel). DMSO and MTT were acquired from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Trypsin-EDTA solution, penicillin-streptomycin solution, mitochondrial membrane potential (MMP) assay kit with JC-1, caspase 3, 8 activity assay kit, and propidium iodide (PI)/RNase staining solution were obtained from Beyotime Biotechnology Co., Ltd. (Shanghai, China). FITC goat anti-rabbit IgG and Annexin V-FITC Apoptosis Detection Kit were acquired from Tianjin Sungene Biotech Co. Ltd. (Tianjin, China). Anti-Bax, anti-Bcl-2, anti-caspase 3, anti-caspase 8, anti-PARP, anti-cyclin D1 (anti-CCND1), anti-cyclin-dependent kinase 4 (anti-CDK4), anti-phosphorylated Rb (anti-p-Rb) (Ser780), and β-actin antibodies were purchased from Bioss Biotechnology Co. Ltd. (Beijing, China).
3
Annexin V-FITC Apoptosis Assay
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Cell apoptosis was detected using Annexin V-FITC Apoptosis Detection Kit (Sungene Biotech Co., Ltd., Tianjin, China) as described in the instruction of manufacture. Briefly, cells inoculated in 6-well plates with 3 × 105 cells each well were suffered with different treatment, and then collected and resuspended in 1 mL of 1 × buffer. 100 μL of cell suspension above was transferred into a new EP tube and incubated with 5 μL of Annexin V-FITC as well as 5 μL of propidium iodide (PI) at room temperature (r.t.) for 5 min in a dark environment. After 500 μL of PBS was dropped, apoptosis of cells was detected by a flow cytometer (BD Accuri™ C6, Piscataway, NJ, USA) and the results were analyzed by FlowJo 7.6 software (Tree Star Inc., Marina Del Rey, CA, USA).
4
Napabucasin-induced Apoptosis in HCC
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HCC cells were seeded in a 6-well plate at the density of 2 × 105 cells per well, cultured overnight, and incubated with napabucasin for an additional 4 or 12 h. The cells were harvested, washed with phosphate buffered saline (PBS), and stained using the Annexin V-FITC Apoptosis Detection Kit (Sungene biotech, Tianjin, China) according to the manufacturer’s instructions. The stained cells were acquired by FACSCalibur flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA), and the percentages of apoptotic cells were analyzed using the FlowJo software (Tree Star, San Carlos, CA, USA).
5
Annexin V-FITC Apoptosis Assay
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An Annexin V-FITC apoptosis detection kit (Sungene Biotech) was used to assess cell apoptosis according to the manufacturer's protocol. Cells were seeded into six-well plates (1×105 cells/well) and cultured until reaching 85% confluence. After digestion, suspension and centrifugation (1,000 × g at 22°C for 3 min), the precipitate was obtained, the cells were resuspended in 300 µl of binding buffer, and 5 µl of Annexin propidium iodide was added. Annexin propidium iodide was mixed and incubated for 10 min in the dark before detection with a BD FACSAriaIII flow cytometer (BD Biosciences).
6
Apoptosis Detection in K562 Cells
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An Annexin V-FITC Apoptosis Detection Kit (Sungene Biotech, Tianjing) was used to detect the percentage of apoptotic cells. K562 and K562/G01 cells (106 cells) were treated for 48 h with KW-2478 at various doses followed by Annexin-V/PI staining. Then, cell apoptosis was detected by flow cytometry (Beckman Coulter, USA).
7
Evaluating Cellular Apoptosis via Flow Cytometry
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For the evaluation of cellular apoptosis, GSK-J4-treated cells were seeded into 6-well plates at a density of 5 × 105 cells/well. After culturing for 48 h, the cells were harvested and washed twice with PBS. The rate of apoptotic cells was determined by the Annexin V-FITC Apoptosis Detection Kit (SUNGENE BIOTECH, Tianjing). Flow cytometric analysis was performed using a CytoFLEX flow cytometer (Beckman Coulter, USA). The apoptotic rate of PKC-α siRNA-transfected cells was evaluated using the same method.
8
Annexin-V-FITC Apoptosis Assay in K562 Cells
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Apoptotic cells were assayed by the Annexin-V-FITC Apoptosis Detection Kit (Tianjin Sungene Biotech Co., Ltd.) according to the manufacturer's instructions. K562 cells were seeded in six-well plates and treated with DMSO or compound 2m (30 nM, 300 nM) for 0, 6, 12, 24 and 48 h respectively. After the treatment, cells were harvested, washed twice with ice-cold PBS at 2500 rpm and resuspended in 1 × Binding buffer. Then, the cells were stained with 5 μl of Annexin-V-FITC for 10 min and 5 μl of PI (50 μg/mL) for 10 min at room temperature in dark and analyzed by flow cytometry. The fraction of cell population in different quadrants was analyzed using quadrant statistics.
9
Annexin V-FITC Apoptosis Assay
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Cell apoptosis was measured with an Annexin V-FITC Apoptosis Detection Kit (Tianjin Sungene Biotech Co., Ltd.). The cells were digested and harvested by trypsin. After being washed twice with PBS, the cells were re-suspended with a 1X binding buffer containing Annexin V-FITC (200 ng/mL). After incubation for 15 min in the dark at room temperature, the cells were washed in PBS and resuspended in binding buffer. PI was added into the cells before flow cytometric analysis. Stained cells were analyzed in FACSCantoTM (BD Biosciences). Data analysis was performed with FlowJo software (Tree Star, Ashland, OR).
10
Monitoring Apoptosis in K562 Cells
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The extent of apoptotic cells were monitored by the Annexin-V-FITC Apoptosis Detection Kit (Tianjin Sungene Biotech Co., Ltd.) on the basis of the manufacturer's instructions. K562 cells (5 × 104 cells per mL) were treated with DMSO or 5-acetamido-1-(methoxybenzyl) isatin (1 μM) for 0, 6, 12, 24 and 48 h respectively. After the exposure, cells were collected, washed with PBS at 461 g and gently resuspended in 1 × binding buffer. Then, the cells were incubated with Annexin-V-FITC (Annexin V-Fluorescein isothiocyanate) and PI (propidium iodide) for 10 min in dark at room temperature and evaluated by flow cytometry. Subsequently, quadrant statistics was used to analyze the fraction of cell population in different quadrants.
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