TUNEL was performed to detect apoptotic cells in mouse tissue slides using a TUNEL kit according to the manufacturer’s instructions (Thermo Fisher Scientific). In brief, 4-μm-thick paraffin sections were deparaffinized in xylene three times for 5 min and hydrated with different concentration of ethanol. Apoptotic cells were stained with diaminobenzidine (DAB) reaction mixture supplied by the kit. The apoptotic cell nuclei were stained in brown and observed under a light microscope (Olympus, Tokyo, Japan). The positive rates were measured using Image J software.
Tunel kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The TUNEL kit is a laboratory assay used for the detection and quantification of apoptosis, a type of programmed cell death. The kit utilizes the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method to label and identify DNA fragmentation, a hallmark of apoptosis. The kit provides the necessary reagents and protocols to perform this analysis.
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Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Nlrp3, by Gentex (2 mentions)
Cd 11b, by Gentex (2 mentions)
Pla goat plus, by Merck Group (2 mentions)
Duolink pla rabbit minus, by Merck Group (2 mentions)
Duolink pla kit, by Merck Group (2 mentions)
Cx31 p, by Olympus (1 mentions)
Inverted fluorescence microscope, by Olympus (1 mentions)
Annexin 5 fitc pi staining kit, by Thermo Fisher Scientific (1 mentions)
A1r hd confocal microscope, by Nikon (1 mentions)
Tcs sp8, by Leica (1 mentions)
Rhodamine 123 rh123, by Merck Group (1 mentions)
Propidium iodide, by Thermo Fisher Scientific (1 mentions)
Annexin 5 fitc, by Thermo Fisher Scientific (1 mentions)
Mitosox kit, by Thermo Fisher Scientific (1 mentions)
Carbonyl cyanide m chlorophenyl hydrazone, by Merck Group (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Nile red, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
19 protocols using tunel kit
1
TUNEL Assay for Apoptosis Detection
2
Lung Tissue Analysis for Apoptosis and NLRP3
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Lung tissue was cut to 4 microns after fixation and embedding. During the experiment, try to ensure that the slices are not contaminated, and at the same time, ensure the storage time of the samples, so that they can be processed in time and follow-up experiments can be carried out in time. According to a previous study [26 (link)], the sections were dewaxed and stained with hematoxylin and eosin to observe lung pathological changes. The sections were stained according to the experimental steps indicated by TUNEL kit (Thermo Fisher Scientific, UA) to observe the death of apoptotic cells. In addition, the expression of NLRP3 in Sample tissues was assessed by staining for NLRP3 using immunohistochemical methods.
3
Apoptosis Detection in Mouse Tissue
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TUNEL was performed to detect apoptotic cells in mouse tissue slides using a TUNEL kit according to the manufacturer’s instructions (Thermo Fisher Scientific).
4
Quantifying Apoptosis in Exosome-Treated HCC Cells
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The TUNEL kit (Thermo Fisher) was performed to identify apoptosis in exosome-treated/untreated HCC cells. After the prescribed treatment according to the manufacturer’s protocol, Huh7 or HA22T cells were fixed by incubation with 4% (w/v) paraformaldehyde for 15 min at 4 °C. Next, the desired cells were sequentially incubated with TUNEL reaction liquid and DAPI for staining. Where DAPI was utilized to stain the nucleus (blue), and TUNEL staining (green) was applied to label apoptotic cells. Finally, fluorescence microscopy (CX31-P, Olympus) was employed to determine the number of TUNEL-positive cells after magnified imaging, and the degree of apoptosis was evaluated.
5
Quantifying Ovarian Cell Apoptosis
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Apoptosis was examined by the TUNEL method. Briefly, ovarian tissues were fixed in 4% paraformaldehyde, embedded in paraffin, cut into sections with a thickness of 5 μm, and incubated with the TUNEL kit (Thermo Fisher Scientific, Waltham, MA, USA) in accordance with the manufacturer’s instructions, followed by visualization under an inverted fluorescence microscope (Olympus Corporation, Tokyo, Japan). The apoptotic index was calculated as the percentage of TUNEL-positive cells in granulosa cells.
6
Apoptosis Measurement: TUNEL and Flow
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The apoptotic profiles of cell and tissue samples were measured using TUNEL and flow cytometry assays, respectively. For flow cytometry assay, an Annexin V‐FITC/PI staining kit (Invitrogen) was used according the kit instructions. For TUNEL assay, a TUNEL kit (Thermo Fisher) was used.
7
Apoptosis Detection in U87 Cells
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Apoptosis was determined with a TUNEL kit (Thermo Fisher Scientific, Shanghai, China) according to the manufacturer’s instructions. Briefly, U87 cells were cultured on coverslips and treated with 4 μM 3-MA or 20 μM CQ for 1 h. At 48 h posttreatment, the cells were fixed with 4% paraformaldehyde (PFA) solution for 10 min at room temperature (37°C). Then in order to block endogenous peroxidase activity, the cells were exposed to 500 μl of methanol with 0.3% H2O2 for 20 min at 37°C. After that, the cells were incubated with TUNEL reaction mixture at 37°C for 60 min and visualized with confocal microscopy (A1; Nikon, Japan).
8
TUNEL Apoptotic Cell Detection
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Apoptotic cells were detected using a TUNEL kit (Thermo Fisher Scientific), according to the manufacturer's instructions. Briefly, paraffin-embedded sections were deparaffinized and hydrated in a graded ethanol series and then digested with trypsin for 40 min at room temperature, and then were incubated with TUNEL reaction buffer in a 37 °C for 1 h, washed with PBS. The apoptotic cells were observed under a light microscope (Olympus, Tokyo, Japan).
9
TUNEL Assay for Apoptosis Quantification
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Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining of HPAECs was performed using a TUNEL kit (Cat# C10617, Thermofisher) as per manufacturer instructions. Five randomly selected fields were photographed, and the number of apoptotic nuclei as well as the total number of nuclei was counted per HPF. The apoptotic index was obtained by means of the formula: number of apoptotic cells per field / total number of cells per field as previously described31 (link).
10
TUNEL Assay for Apoptosis Detection
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A TUNEL kit (C10617, Life technologies) was used to detect apoptotic cells according to the manufacturer’s protocols. Samples were prepared per paraffin protocol outlined above and then incubated with TUNEL working solution for 1 h and shielded from light at 37 °C. The nuclei were stained by Dapi for 30 min. The specimens were imaged using a Nikon A1R HD confocal microscope with a DU4G filter-based detector, using a Super Plan Fluor LWD 20x 0.70NA air objective lens with digital zoom of 1x.
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