Cell proliferation of INS-1E cells was determined by means of a colorimetric MTT-assay as previously reported [26 (link)]. Briefly, INS-1E cells were serum-deprived overnight and exposed to leptin or/and TNF-α for 48 h. Then, the cells were incubated with 20 μl 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT; Sigma-Aldrich, St. Louis, MO, USA) at 37°C for 3 h. After removal of MTT, 150 μl DMSO was added to the cells and the absorbance was measured at 492 nm with a microplate reader. Apoptosis assay was performed with Annexin V-FITC apoptosis assay kit (Bender MedSystems) in accordance with the manufacturer's directions.
Annexin 5 fitc apoptosis assay kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Annexin V-FITC apoptosis assay kit is a tool for the detection and quantification of apoptosis in cell populations. It utilizes Annexin V, a protein that binds to phosphatidylserine, a phospholipid that is translocated to the outer membrane of apoptotic cells. The kit also includes FITC, a fluorescent label, which allows for the visualization and analysis of apoptotic cells using flow cytometry or fluorescence microscopy.
Automatically generated - may contain errors
Lab products found in correlation
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Tetrahydrouridine, by Merck Group (1 mentions)
Squalene, by Merck Group (1 mentions)
Colchicine, by Merck Group (1 mentions)
2 deoxycytidine, by Merck Group (1 mentions)
Cell culture reagents, by Thermo Fisher Scientific (1 mentions)
Sodium acetate, by Merck Group (1 mentions)
Menthol, by Merck Group (1 mentions)
3 4 dimethylthiazol 2 yl 2 5 diphenyl tetrazolium bromide mtt, by Thermo Fisher Scientific (1 mentions)
Rnase a, by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Flow cytometry, by BD (1 mentions)
6 protocols using annexin 5 fitc apoptosis assay kit
1
Measuring INS-1E Cell Proliferation and Apoptosis
2
PEDF Induces Apoptosis in HemEC
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HemEC were seeded at 5 × 104 per well in 6‐well plates and were allowed to attach overnight. The cell cultures were treated with PEDF at different concentrations for 48 hours. Cells were harvested according to the manufacturer's recommendations. Flow cytometry was carried out to measure apoptosis using the Annexin V‐FITC Apoptosis Assay Kit (Bender Med Systems, Vienna, Austria). Cells treated with 25 μmol/L colchicine were used as positive controls, and cells treated with PBS were used as negative controls.
3
Apoptosis and Cell Cycle Analysis
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The flow cytometry was adopted to determine cell apoptosis and cell cycle specifically as follows: The cells transfected for 48 h were prepared into cell suspension with 1*106 cells, and the suspension was transferred to a cell culture flask for growth overnight. The collected cells were washed with PBS, and their apoptosis rate was evaluated by an Annexin V-FITC Apoptosis Assay Kit (Invitrogen™, the United States) according to the kit instructions.
4
Synthesis and Characterization of Anticancer Compounds
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Gemcitabine was purchased from Sequoia Research Product, Ltd. (UK). Squalene, 3-[4,5-dimethylthiazol-2-yl]-3,5-diphenyl tetrazolium bromide (MTT), 2′-deoxycytidine, colchicine and sodium acetate were purchased from Sigma-Aldrich Chemical Co. (St. Quentin Fallavier, France). Tetrahydrouridine was purchased from Merck Chemicals, Ltd. (Nottingham, UK). All solvents were purchased from Carlo Erba (Val-de-Reuil, France). CholEsteryl BODIPY 542/563, CholEsteryl BODIPY FL, Annexin V-FITC apoptosis assay kit and cell culture reagents were purchased from Invitrogen-Life Technologies (Cergy Pontoise, France). The synthesis of squalenoyl-gemcitabine (SQ-gem), 1,1′,2-tris-nor-squalenic acid and diarylethylene isoCA-4 has been previously reported by us (Figure 1 ).21 (link),27 (link)
5
Menthol-Induced Apoptosis and Autophagy
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Menthol (Sigma-Aldrich, Schnelldorf, Germany, Cat No. 63660) was purchased from Sigma Aldrich (St. Louis, CA, USA) and dissolved in DMSO for storage at 4 °C. Moreover, RNase A and propidium iodide (PI) were purchased from Sigma Aldrich (St. Louis, CA, USA). 3-(4-dimethylthiazol-2-yl)-2-5-diphenyl tetrazolium bromide (MTT) and Annexin V-FITC apoptosis assay kit was obtained from Invitrogen (Walthem, MA, USA) and BD Bioscience (Palo Alto, CA, USA), respectively. Autophagy LC3-II antibody-based assay kit was purchased from Luminex Corporation (Austin, TX, USA). All other reagents used in the experiments were commercially analytical grade and available.
6
Quantifying Apoptosis in NSCLC Cells
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Apoptosis was determined using the Annexin V-FITC apoptosis assay kit (Cat No.
BMS500FI-300, Invitrogen, Carlsbad, CA, USA). A549 and H460 cells were trypsinized and
rinsed with cold PBS. Subsequently, NSCLC cells (1×105 cells) were resuspended
in 100 μL of binding buffer. After 20 minutes of incubation with 5 μL of Propidium iodide
(PI) and 5 μL of Annexin V-FITC, apoptotic cells were immediately analyzed by a flow
cytometry (BD Biosciences, San Jose, CA, USA).
BMS500FI-300, Invitrogen, Carlsbad, CA, USA). A549 and H460 cells were trypsinized and
rinsed with cold PBS. Subsequently, NSCLC cells (1×105 cells) were resuspended
in 100 μL of binding buffer. After 20 minutes of incubation with 5 μL of Propidium iodide
(PI) and 5 μL of Annexin V-FITC, apoptotic cells were immediately analyzed by a flow
cytometry (BD Biosciences, San Jose, CA, USA).
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