Chondrocyte apoptosis after transfection was determined by TUNEL assay using a kit (C1086; Beyotime Biotechnology, Jiangsu, China) according to the manufacturer’s instructions. Specimens were visualized under a confocal microscope (Leica). Chondrocytes after transfection were also detected by Annexin V-FITC/propidium iodide double staining (V13241; Life Technologies) with FACS analysis.
V13241
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
The V13241 is a laboratory instrument manufactured by Thermo Fisher Scientific. It is designed for precision temperature control and monitoring applications within a laboratory setting. The core function of the V13241 is to provide accurate and reliable temperature regulation for a variety of experimental and analytical processes.
Automatically generated - may contain errors
Lab products found in correlation
P3566, by Thermo Fisher Scientific (2 mentions)
A1r confocal microscope, by Nikon (2 mentions)
C1086, by Beyotime (1 mentions)
Ra 6023, by Cell Biologics (1 mentions)
As 60804, by AnaSpec (1 mentions)
Flow cytometry, by BD (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Cck 8 assay, by Dojindo Laboratories (1 mentions)
Nis element, by Nikon (1 mentions)
Tie fluorescent microscope, by Nikon (1 mentions)
Spectra x, by Lumencor (1 mentions)
Alamarblue assay, by Thermo Fisher Scientific (1 mentions)
Annexin 5, by Thermo Fisher Scientific (1 mentions)
Fix perm kit, by BD (1 mentions)
Click it edu reaction, by Thermo Fisher Scientific (1 mentions)
19 protocols using v13241
1
Chondrocyte Apoptosis Assessment
2
Amylin-induced Endothelial Cell Apoptosis
3
Cell Cycle and Apoptosis Analysis
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Cells were collected and resuspended in Tris-buffered saline with 10 µg/mL DAPI and 0.1% nonidet P-40 detergent for flow cytometry. Cell cycle phases were analyzed using FlowJo software. Apoptosis was measured using FITC-Annexin V antibody and PI staining following manufacturer’s instruction (Life Technology, V13241). Cells were read using flow cytometry (BD Bioscience), and the results were analyzed via FlowJo.
4
Apoptosis Assay in HeLa Cells
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HeLa cells were seeded in a 25-mL T-flask (Thermo Scientific) in DMEM (Life Technologies) supplemented with 10% FBS and 1% penicillin/streptomycin at 37 °C under 5% CO2. The cells were then incubated with the peptides at 10 μM for a period of 1 h, 4 h, 6 h, 8 h, and 10 h. Similarly, self-assembled peptides at 100 μM were incubated for a period of 3 h in PBS solution in triplicate. The cells were collected and washed in cold PBS. Following the manufacturer’s protocol (Life Technologies, V13241), the cells were incubated with 5 µl of Alexa Fluor 488-conjugated annexin V and 1 µl of 100 µg ml–1 PI working solution in 100 µl of annexin-binding buffer solution for 15 min at room temperature. After the incubation period, 400 µl of 1× annexin-binding buffer was added with gentle mixing, and the samples were kept on ice. The stained cells were then analyzed by flow cytometry (FACS ealibur, BD Bioscience), with the fluorescence emission measured at 530 nm (i.e., FL1) and >575 nm (i.e., FL3).
5
AdipoRon Induces Apoptosis in Cell Lines
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Cells were dissociated with trypsin, counted, and 100,000 Cells were seeded in each well of a 24 well plate and allowed to adhere in full media overnight. The next day, media was replaced with treatment media consisting of DMEM supplemented with 2.5% FBS and either DMSO or AdipoRon (50μM) and incubated for an additional 24h. Annexin V staining was performed following the manufacturer’s protocol (Life Technologies, V13241). Cells were assessed by flow cytometry comparing propidium iodide versus Annexin V positive cells.
6
Probing Cellular Mechanoresponse in 3D Collagen
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Collagen gels embedded with either vim+/+ or vim−/− mEFs (400,000 cells/ml; see above) were prepared in culture dishes such that the dimensions of the gel were 20-mm in diameter and 1-mm in height. After 24 h, the gels were deposited onto a Kinexus rheometer (Malvern) equipped with a 20-mm circular parallel plate geometry and Peltier plate that maintains the sample temperature at 37°C. The upper plate was lowered to contact the gel. Cell culture medium was added around the sample to prevent drying and allow free fluid flow. The gel was allowed to relax for a minimum of 3 min between the plates and then subjected to a step compression of 25%, 50%, or 80% strain, which was held for 3 min to allow for axial stresses to relax to steady state. The plate was raised, and the gel was transferred to a culture dish with cell culture medium. Cell fate was determined in 3D collagen gels by staining with propidium iodide (Invitrogen, V13241) according to the manufacturer’s instructions. The cell membrane is impermeable to propidium iodide, and positive staining indicates rupture of the cell membrane and necrotic cell death. Cells were imaged at multiple locations throughout the gel with a 10× objective and manually counted in 800 × 800-µm2 areas (five to eight locations per condition). Experiments were conducted a minimum of two times, yielding 300–700 cells per condition.
7
Time-lapse Imaging of Alu-Induced Apoptosis
8
Apoptosis Detection Using Invitrogen Kit
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Cell apoptosis was detected by using a kit to the product manual (Invitrogen, V13241).
9
Apoptosis Quantification in NK Cells
10
Annexin V-FITC Apoptosis Detection
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Cell death was determined using annexin V/propidium iodide (PI) staining as directed by the manufacturer (Invitrogen no. V13241). Huh7 cells were infected with the serotypes of dengue at 1 MOI, and 60 h postinfection the cells were trypsinized and collected in 1× cold PBS. After cell density was determined, about 1 × 106 cells were centrifuged and resuspended in 100 μL of 1× annexin V binding buffer. The cell suspension was then incubated for 15 min at room temperature with 5 μL Alexa Fluor 488 annexin V (component A) and 1 μL 100 μg/mL PI working solution. Following incubation, 400 μL of 1× annexin V binding buffer was gently mixed into the cell suspension, and the mixture was immediately placed on ice. The stained cells were analyzed using a FACSCalibur device (Beckton and Dickinson), and the data were evaluated using FlowJo software.
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