Dihydromyricetin (DMY) was purchased from Chengdu Kangbang Biotechnology, sodium nitroprusside, MTT, DMSO, DHE and Annexin V‐FITC/PI were purchased from Sigma‐Aldrich; Endothelial cell medium was from ScienCell; Hoechst 33342 were from Beyotime Institute of Biotechnology. PI3K inhibitor LY294002 was from Calbiochem (La Jolla, CA). And the various antibodies used in the experiments were obtained from Cell Signaling Technology.
Annexin 5 fitc pi
Manufactured by Merck Group
Sourced in United States, Germany
Annexin V-FITC/PI is a laboratory reagent used in flow cytometry applications. It contains Annexin V, a protein that binds to phosphatidylserine, and propidium iodide (PI), a DNA-binding dye. This combination allows for the detection and quantification of apoptotic and necrotic cells.
Automatically generated - may contain errors
Lab products found in correlation
Propidium iodide, by Merck Group (3 mentions)
Facscalibur, by BD (3 mentions)
Facscalibur flow cytometer, by BD (2 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Sodium nitroprusside, by Merck Group (1 mentions)
Hoechst 33342, by Beyotime (1 mentions)
Endothelial cell medium (ecm), by ScienCell (1 mentions)
Np 40, by Merck Group (1 mentions)
Pristimerin, by Merck Group (1 mentions)
Akt sirna, by RiboBio (1 mentions)
Cell counting kit 8 (cck8), by Merck Group (1 mentions)
Muse cell analyser, by Merck Group (1 mentions)
Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions)
Acea flow cytometer, by Agilent Technologies (1 mentions)
Annexin 5 fitc propidium iodide staining, by BD (1 mentions)
Novoexpress version 1, by Agilent Technologies (1 mentions)
15 protocols using annexin 5 fitc pi
1
Dihydromyricetin-mediated Vascular Protection
2
Cell Cycle and Apoptosis Analysis
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For the cell cycle analysis, the cells were fixed overnight in chilled methanol before staining with 50 μg/mL propidium iodide (PI, Sigma-Aldrich) in the presence of 1 mg/mL RNase (100 units/mL; Sigma-Aldrich) and 0.1% NP40 (Sigma-Aldrich).
For the apoptosis analysis, the samples were incubated with Annexin V- FITC/PI according to the manufacturer's recommended protocol (Sigma-Aldrich). Cell-bound fluorescence was analyzed using a FACSCalibur flow cytometer (Becton Dickinson, CA, USA).
For the apoptosis analysis, the samples were incubated with Annexin V- FITC/PI according to the manufacturer's recommended protocol (Sigma-Aldrich). Cell-bound fluorescence was analyzed using a FACSCalibur flow cytometer (Becton Dickinson, CA, USA).
3
Apoptosis Quantification in HaCaT Cells
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Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double staining was used to measure percentile of apoptosis in HaCaT cells. Cells at a concentration of 1 × 105 cells/mL were seeded in 6-well plates. Finally, the cells were resuspended in 500 μL of 1x binding buffer and mixed with Annexin V-FITC/PI (Cat number APOAF, Sigma Chemical Co., St. Louis, MO, USA). After incubation for 30 min, the cells were measured by Accuri C6 flow cytometry (Accuri, Ann Arbor, MI, USA).
4
Flow Cytometric Analysis of Cell Apoptosis
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Transfected cells of SPC-A1 and A549 were harvested and re-suspended with binding buffer at the density of 1 × 106 cells/ml. Subsequently, Annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI; Sigma, St. Louis, MO, U.S.A.) was performed to stain cells according to manufacturer’s introductions. Cell apoptotic rate was analyzed via a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, U.S.A.).
5
Cell Cycle and Apoptosis Analysis
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For the cell cycle analysis, the cells were stained with propidium iodide (PI, Sigma‐Aldrich). The apoptosis analysis was performed using Annexin V‐FITC/PI according to the manufacturer's protocol (Sigma‐Aldrich).
6
Pristimerin-Induced Apoptosis Pathway
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Pristimerin, Annexin V‐FITC/PI, JC‐1 were purchased from Sigma‐Aldrich. Akt‐siRNA was obtained from Ribobio. Plasmid for FoxO3a, FoxO3a siRNA and Empty plasmid vectors N1 were kindly provided by Marten P. Smidt (Rudolf Magnus Institute of Neuroscience).
7
Cell Proliferation and Apoptosis Assay
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The Cell Counting Kit-8 (CCK-8) method (Sigma-Aldrich, St. Louis, MO, USA) was used to measure cell proliferation. The analysis of the cell cycle and apoptosis was detected using a FACSCalibur flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA). For the cell cycle analysis, propidium iodide (PI, Sigma-Aldrich) was used to stain the cells. According to the manufacturer's instructions, Annexin V-FITC /PI (Sigma-Aldrich) was used for apoptotic analysis. The CCK-8 and flow cytometry (FCM) assays were performed as previously described [33 (link)].
8
Apoptosis Assessment via Annexin V-FITC/PI
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Approximately 1.5–2×105 cells per well were plated in six-well plates, treated with 10–20 mM PF for 24 hours or 20 mM for 6, 12, and 24 hours, then collected and stained with Annexin V-FITC/PI according to the manufacturer’s instructions (Sigma-Aldrich). Briefly, the cells were washed twice with cold phosphate-buffered saline (PBS), resuspended in binding buffer at 5×105 cells/500 μL, stained with 5 μL Annexin V and 10 μL Propidium iodide (PI), and incubated for 15 minutes at room temperature in the dark prior to flow cytometric analysis.
9
Apoptosis Assay for H1299 Cells
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The LUAD cells were seeded into 6-well plates and cultured in a saturated humidity incubator containing 5% CO2 at 37°C for 48 hours. Trypsin was used to digest the cells, and the serum-containing medium was used to terminate digestion and resuspend cells. The cells were centrifuged at 4°C for 10 minutes at 1000 r/min, and the supernatant was discarded. Then, 1 mL PBS was used to resuspend cells and cells were centrifuged for 10 minutes to discard the supernatant. After the cells were washed twice, the apoptosis rate of H1299 cells was tested using Annexin V-FITC/PI (Merck Sigma-Aldrich, Germany) by FCM (Thermo Fisher Scientific).
10
Quantifying Apoptosis in HaCaT Cells
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Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double staining was used to measure percentile of apoptosis in HaCaT cells. After 24 h later with UVB irradiation and sample treatment, the cells were re-suspended in 500 μl of 1× binding buffer and mixed with Annexin V-FITC/PI (Cat number APOAF, Sigma, United States). After incubation for 30 min, the cells were measured by Accuri C6 flow cytometry (Accuri, Ann Arbor, United States).
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