Using the TUNEL (terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling) method, lung sections were stained for the detection of fragmented DNA, which is indicative of apoptotic cells. Sections were incubated with TdT-reaction solution and nuclei were visualized using TUNEL reagents in a kit (Promega, Madison, WI) and DAPI nuclear stain. Fluorescence images were obtained on a Nikon E100 microscope. Quantification of TUNEL-positive cells was by determining the percentage of TUNEL-positive (red) cells in multiple high power fields (n = 3 sections per mouse strain and treatment group). Data are reported as mean percentage of TUNEL-positive cells per lung section ± SEM. Data are reported as mean percentage of positive cells per lung section ± SEM.
Tunel reagent
Manufactured by Promega
Sourced in United States
The TUNEL reagent is a laboratory tool used to detect and quantify apoptosis, a form of programmed cell death. It works by labeling the DNA fragments generated during apoptosis, allowing for their visualization and analysis. The TUNEL reagent is a versatile tool for researchers studying cell biology, developmental processes, and disease mechanisms.
Automatically generated - may contain errors
Lab products found in correlation
E100 microscope, by Nikon (1 mentions)
Annexin 5 fitc, by BD (1 mentions)
Alexa fluor 488 conjugated goat anti mouse igg, by Cell Signaling Technology (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Bx61 fluorescence microscope, by Olympus (1 mentions)
510 meta laser scanning confocal microscope, by Zeiss (1 mentions)
Mounting medium, by Beyotime (1 mentions)
6 protocols using tunel reagent
1
Quantification of Apoptosis in Lung Tissue
2
TUNEL Assay for Apoptosis Analysis
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By virtue of the TUNEL approach, heart slices were dyed for identifying DNA fragmentation, which could reflect cell apoptosis. Slices were cultivated in TdT-reaction liquor and the visualization of nuclei was realized via TUNEL reagents (Promega, America) and DAPI nuclear dye. Fluorescent pictures were captured via a Nikon microscopic device. Quantitation of TUNEL-positive cells was completed via identifying the corresponding proportion (%) (green) in several high-power fields (n = 3 slices every mouse strain and treatment group).
3
Peptide Modulation of Apoptosis in HeLa Cells
4
TUNEL Assay for Apoptosis in MSCs
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After treatment with STS for 16 h, MSCs were fixed with 4% PFA and permeabilized. According to the manufacture's protocol, cells were incubated with TUNEL reagent (G3250, Promega) for 60 min at 37 °C. Positive cells were counted after counterstained with DAPI under microscope.
5
In Situ Apoptosis Detection Using TUNEL Assay
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The terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) assay was used for in situ apoptosis with TUNEL reagent (Promega, Madison, Wisconsin, USA). In brief, 10 μm paraffin sections were treated with proteinase K 20 μg/mL and then incubated in a nucleotide mixture containing fluorescein-12-dUTP and TdT. Positive controls were pretreated with 1 U/mL DNases, and negative controls were incubated without TdT. Dark yellow apoptotic cells were counted under light microscopy (CX31, Olympus, Japan).
6
Cell Proliferation and Apoptosis Analysis
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Immunofluorescence staining of Ki67 and TUNEL were performed to detect cell proliferation and apoptosis, respectively. Cells cultured on coverslips in 35-mm culture dishes were treated with PPD to ensure a discrimination of individual nuclear foci in immunofluorescence staining. After 24 h incubation, cells were fixed with 4% paraform aldehyde for 20 min at room temperature and permeabilized with 0.1% Triton X-100 (Sigma, Louis, MO, USA) for 10 min at 4 °C. After blocking with 5% Bovine Serum Albumin for 30 min at room temperature, cells were stained with TUNEL reagent (Promega, Madison, WI, USA) for 60 min at room temperature in the dark for in situ apoptosis detection; or incubated with antibody against Ki67 (EMD Millipore, Beijing, China) overnight at 4 °C, followed by incubated with Alexa Fluor 488-conjugated goat anti-mouse IgG (Cell Signaling Technology, Danvers, MA, USA) for 1 h at room temperature. Thereafter, samples were counterstained with a mounting medium (Beyotime, Shanghai, China) containing Hoechst 33,342. Three random fields of cell samples were inspected with an Olympus BX61 fluorescence microscope (Olympus, Tokyo, Japan), and images were captured on a Zeiss 510 Meta laser scanning confocal microscope (Zeiss, Jena, Germany). Nuclei containing ≥10 immunoreactive foci were scored as positive for TUNEL.
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