An Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) kit (BD Biosciences, NJ, USA) was used to examine cell apoptosis. Briefly, transfected KYSE-150 and TE-1 cells were seeded in 6-well plates. The cells were suspended in binding buffer and labeled with Annexin V-FITC and PI for 15 min in a dark environment. The labeled cells were analyzed on the flow cytometer (BD Biosciences). The apoptosis rate of cells was examined by Flow Jo program (NIH, Bethesda, MD, USA) [17 (link)].
Annexin 5 fluorescein isothiocyanate fitc propidium iodide kit
Manufactured by BD
Sourced in United States
The Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) kit is a laboratory reagent used for the identification and quantification of apoptotic and necrotic cells. The kit utilizes Annexin V, a calcium-dependent phospholipid-binding protein, conjugated to the fluorescent dye FITC, and the nuclear stain propidium iodide. This combination allows for the discrimination between early apoptotic, late apoptotic, and necrotic cells through flow cytometric analysis.
Automatically generated - may contain errors
Lab products found in correlation
Cellquest software version 0, by BD (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Bcl 2, by Santa Cruz Biotechnology (2 mentions)
Cleaved caspase 3, by Santa Cruz Biotechnology (2 mentions)
Facsdiva software, by BD (2 mentions)
Flow cytometry, by BD (2 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (2 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Bcl2 associated x (bax), by Santa Cruz Biotechnology (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions)
Modfit lt 3, by Verity Software House (1 mentions)
Guava easycyte ht, by Merck Group (1 mentions)
Cytoflex, by Beckman Coulter (1 mentions)
Facsaria, by BD (1 mentions)
Facscalibur, by BD (1 mentions)
Transwell chamber, by BD (1 mentions)
Vascular endothelial growth factor (vegf), by Santa Cruz Biotechnology (1 mentions)
23 protocols using annexin 5 fluorescein isothiocyanate fitc propidium iodide kit
1
Annexin V-FITC/PI Apoptosis Assay
2
Puerarin Attenuates Oxidative Stress and Inflammation
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Puerarin (PEU, purity >99.5%), dimethyl sulfoxide (DMSO), 2,7-dichorofluorescin diacetate (DCFH-DA), and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were obtained from Sigma (St. Louis, MO, U.S.A.). Dulbecco’s modified Eagle medium (DMEM) and fetalbovine serum (FBS) were purchased from Life Technologies Inc (Gaithersburg,MD, USA). The levels of interleukin 1 beta (IL-1β), interleukin-6 (IL-6) and cytokines IL-10 commercially available enzyme-linked immunosorbent assay (ELISA) kits were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Primary antibodies against Bcl-2, cleaved-caspase-3, Keap1, Bax, Nrf2, HO-1, NQO1and β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). BCA protein assay kit, Horseradish peroxidase (HRP) conjugated secondary antibody and enhanced chemiluminescence (ECL) reagent were from Pierce Chemical (Rockford, IL, USA). Annexin V‑fluorescein isothiocyanate (FITC)/propidium iodide (PI) kit and the lactate dehydrogenase (LDH) assay kit were from BD Biosciences (San Jose, CA, USA). All other reagents used in this experiment were of analytical grade commercially available.
3
Apoptosis and Cell Cycle Analysis
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For apoptosis detection, an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) kit (BD Biosciences) was used according to the manufacturer’s instructions. Briefly, the transfected cells were first incubated with FITC-conjugated Annexin V at 22°C in the dark for 15 min, and then with PI for 10 min. The samples were analyzed on a flow cytometer (Guava easyCyte HT, EMD Millipore, Billerica, MA), and the apoptosis rate of cells was evaluated using ModFit LT 3.3 software (Verity Software House, Topsham, ME, USA).
For cell cycle detection, transfected cells were sorted in 6-mm culture dishes for 72 h and then cleared in 70% ethanol for 1 h. Thereafter, the cells were washed and incubated with staining buffer (containing 10 µg/mL RNase and PI, Solarbio) at room temperature for 10 min. The cell cycle distribution was determined using the flow cytometer and ModFit LT 3.3 software.
For cell cycle detection, transfected cells were sorted in 6-mm culture dishes for 72 h and then cleared in 70% ethanol for 1 h. Thereafter, the cells were washed and incubated with staining buffer (containing 10 µg/mL RNase and PI, Solarbio) at room temperature for 10 min. The cell cycle distribution was determined using the flow cytometer and ModFit LT 3.3 software.
4
Quantification of Apoptosis by Flow Cytometry
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Apoptosis was quantitated by flow cytometry using an annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) kit (BD Bioscience, San Jose, CA, USA) according to the manufacturer’s instructions. At 48 h post-treatment with a PRMTs type I inhibitor (MS023), cells were washed with phosphate buffered saline (PBS), suspended in annexin V binding buffer and annexin V-FITC solution, and added with PI. These cells were then incubated at RT for 15 min. Stained cells were analyzed using a fluorescence-activated cell sorter (FACS) (BD FACSVesrse™; BD Biosciences). Data were analyzed using FACSDiva™ software (BD Biosciences).
5
Apoptosis Evaluation in H9c2 Cells
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The apoptotic rate of the H9c2 cells was evaluated by flow cytometry using an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) kit (BD Biosciences). Cells in 6-well plates were trypsinized and washed twice in ice-cold PBS. Cells were suspended in binding buffer and 5 μl Annexin V-FITC and 5 μl PI were added to each sample. The cells were incubated at room temperature in the dark for 15 min and analyzed using a flow cytometer (BD Biosciences).
6
Granulosa Cell Apoptosis Assay
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Cell apoptosis was determined by flow cytometry (BD Biosciences, U.S.A.) using an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) kit (BD Biosciences, U.S.A.), according to the manufacturer’s protocol. Briefly, Granulosa cells were seeded in 12-well plates with a density of 1 × 106 cells/well for 24 h and then treated with 3-NPA at 5.0 mmol/l for another 24 h. Granulosa cells were harvested and washed twice with cold PBS, and labeled with Annexin V-FITC and PI for 15 min at 25°C in the dark in binding buffer. Fluorescence intensity of granulosa cells was detected by flow cytometry within 1 h. The analysis was replicated thrice and the apoptosis rate (%) for each treatment was obtained.
7
Annexin V-FITC/PI Apoptosis Assay
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Cells were collected by digesting with pancreatin and centrifuging. After that, cells were re-suspended with 1 × binding buffer after washed with iced phosphate-buffered saline (PBS). Later, these cells were processed following the manufacturer's instructions of Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) kit (BD Pharmingen, San Diego, CA, USA). Apoptotic cells were examined using flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA), and then apoptosis rate was calculated.
8
Apoptosis Detection in INS-1-3 Cells
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The INS-1-3 cell apoptosis was detected by flow cytometry analysis using an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) kit (BD Pharmingen™, United States). Briefly, INS-1-3 cells seeded in 6-well plates (2×105 cells/well) were incubated for 24 hours with and without palmitic acid (0.3mM) plus glucose(16.7mM). At the end of treatment, cells were harvested by trypsin-EDTA digestion. After washing with PBS, cells were re-suspended in binding buffer and incubated with 5 μL of Annexin V-FITC and PI solution for 15 min at room temperature in the dark. They were analyzed by flow cytometry (Beckman Coulter, Fullerton, CA, USA).
9
Apoptosis Detection by Flow Cytometry
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Cells were detached with 0.25% ethylene diamine tetraacetic acid-depleted trypsin and collected after centrifugation. Cells were resuspended in 1 × binding buffer and detected by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) kit (BD Pharmingen, San Diego, USA) following the manufacturer's instructions. Apoptotic cells were tested by a flow cytometer (BD Biosciences, NJ, USA), and the apoptosis rate was then calculated [26 (link)].
10
Quantifying Apoptosis in Infected Cells
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The apoptosis rate of infected CC cells was detected by flow cytometry analysis. The infected CC cells were stained using an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) kit (BD Biosciences, USA), and the results were calculated with a flow cytometer using the CellQuest software version 0.9.3.1 (BD Biosciences, USA).
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