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Tunel assay

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The TUNEL assay is a method used to detect and quantify apoptosis, or programmed cell death, in cells. It works by labeling the terminal end of nucleic acids that have been fragmented as a result of apoptotic processes. This allows for the identification and visualization of apoptotic cells.

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Lab products found in correlation

Fluorescence microscope, by Olympus (4 mentions) Eclipse e600 microscope, by Nikon (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Wuhan Servicebio Technology (3 mentions) Cleaved caspase 3, by Cell Signaling Technology (2 mentions) Cell counting kit 8 (cck8), by Dojindo Laboratories (2 mentions) C1086, by Beyotime (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Mitotracker red, by Thermo Fisher Scientific (1 mentions) Ab154179, by Abcam (1 mentions) Ab192890, by Abcam (1 mentions) Ab109012, by Abcam (1 mentions) Superoxide dismutase activity assay kit, by Abcam (1 mentions) Tempol, by Merck Group (1 mentions) Annexin 5 fitc apoptosis detection kit, by Thermo Fisher Scientific (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Mitotracker green, by Thermo Fisher Scientific (1 mentions) Mitosox red, by Thermo Fisher Scientific (1 mentions) Mda assay kit, by Beyotime (1 mentions) Dcfh da, by Beyotime (1 mentions)

83 protocols using tunel assay

1

Apoptosis Assessment in Murine Tumors

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Cell apoptosis in mice tumor tissues was examined using TUNEL assay (Biyuntian, Wuxi, China) according to the manufacturer’s instructions.
Li Z., Zhang L., Gao M., Han M., Liu K., Zhang Z., Gong Z., Xing L., Shi X., Lu K, & Gao H. (2019). Endoplasmic reticulum stress triggers Xanthoangelol-induced protective autophagy via activation of JNK/c-Jun Axis in hepatocellular carcinoma. Journal of Experimental & Clinical Cancer Research : CR, 38, 8.
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2

Measuring Late Apoptosis in H9c2 Cells

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The parameters of late apoptosis were measured using a TUNEL assay (Biyuntian, Shanghai, China). At 72 h post-transfection, H9c2 cells were fixed with 4% formaldehyde solution for 25 min at 4°C, rinsed three times with PBS at room temperature, and treated with 0.2% Triton X-100. After incubation buffer was added cells were incubated for 60 min at 37°C. Then, cells were immersed with 3 ml of 2 × sodium chloride-sodium citrate buffer. The reaction was terminated 15 min later. Unbound fluorescein-12-dUTP was washed away with PBS. After addition of 3 ml of propidium iodide (1 μg/ml) was added for 5 min later, and cells were counterstained with hematoxylin and eosin. Finally, TUNEL-positive cells were counted using a microscope.
Gong L., Jiang H., Qiu G., & Sun K. (2021). miR-208a Promotes Apoptosis in H9c2 Cardiomyocytes by Targeting GATA4. Congenital Heart Disease, 16(5), 499-512.
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3

Assessing Hepatocyte Apoptosis in Mice

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Mice were sacrificed, then livers were dissected. After overnight incubation in 4% paraformaldehyde, livers were dehydrated and embedded in paraffin. Sections of 5 µm thickness were cut and mounted on poly-lysine-coated slides, and hepatocyte apoptosis was detected using a TUNEL assay (Beyotime Institute of Biotechnology) according to the instructions of the manufacturer.
Song X., Shen Y., Lao Y., Tao Z., Zeng J., Wang J, & Wu H. (2019). CXCL9 regulates acetaminophen-induced liver injury via CXCR3. Experimental and Therapeutic Medicine, 18(6), 4845-4851.
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4

Oxidative Stress-Induced Apoptosis Assays

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Annexin V-FITC Apoptosis Detection Kit (Invitrogen, China), Ammonium tetrathiomolybdate (TTM) (Sigma-Aldrich, USA), TUNEL assay (Beyotime, China), ROS Assay kit (Beyotime, China), ROS Fluorescent Probe Kit (KeyGEN, China), MDA Assay Kit (Beyotime, China), GSH and GSSG Assay Kit (Beyotime, China), Tempol (Sigma-Aldrich, USA), Cell Counting Kit-8 (Dojindo, Japan), DCFH-DA (Beyotime, China), MitoSOX Red (Invitrogen, China), JC-1 (Invitrogen, China), Mito-Tracker Red, Mito-Tracker Green and Lysosome-Tracker Red (Invitrogen, China), Cell mitochondria isolation kit (Beyotime, China), Superoxide Dismutase Activity Assay kit (ab65354; Abcam).
Antibodies were from various sources, including GAPDH (5174; Cell Signaling Technology), Atox1 (22641-1-AP; Proteintech), Atox1 (ab154179; Abcam), NeuN (ab177487; Abcam), Cleaved Caspase-3 (9661; Cell Signaling Technology), Bax (ab32503; Abcam), Bcl-2 (ab182858; Abcam), DJ-1 (ab76008; Abcam), TOMM20 (ab283317, Abcam), TOMM20 (ab186735, Abcam), LC3-Ⅱ (ab192890, Abcam), SQSTM1/p62 (ab109012, Abcam), PINK1 (23274-1-AP, Proteintech), PRKN (14060-1-AP, Proteintech), COX4 (11242-1-AP, Proteintech), FLAG (ab205606; Abcam).
Zhao P., Shi W., Ye Y., Xu K., Hu J., Chao H., Tao Z., Xu L., Gu W., Zhang L., Wang T., Wang X, & Ji J. (2024). Atox1 protects hippocampal neurons after traumatic brain injury via DJ-1 mediated anti-oxidative stress and mitophagy. Redox Biology, 72, 103156.
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5

Rat Kidney Histology and Apoptosis

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The rat’s kidney was collected and fixed in 10% formalin. Next, the paraffin sections were dewaxed and hydrated; and then slices were incubated in HE solution for 5–20 min. Finally, the kidney slices were observed in a light microscope (Olympus, Tokyo, Japan), and kidney damage was assessed according to previous research. The apoptosis was detected by TUNEL assay based on protocols (Beyotime, Shanghai, China).
Sheng X., Yang Y., Liu J., Yu J., Guo Q., Guan W, & Liu F. (2021). Ophiopogonin A Alleviates Hemorrhagic Shock-Induced Renal Injury via Induction of Nrf2 Expression. Frontiers in Physiology, 11, 619740.
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6

TUNEL Assay for Apoptosis in BALF and Lung

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The cell-debris pellets of bronchoalveolar lavage fluid (BALF) samples and lung tissue (paraffin sections) were prepared to determine apoptosis activity using the TUNEL assay (Beyotime Co., Ltd., Shanghai, China) according to manufacturer's instructions.
Ma Z., Tang X., Gao Y., Wang H., Yu P, & Liu R. (2020). Combined Extracts of Epimedii Folium and Ligustri Lucidi Fructus with Budesonide Attenuate Airway Remodeling in the Asthmatic Rats by Regulating Apoptosis and Autophagy. Evidence-based Complementary and Alternative Medicine : eCAM, 2020, 2319409.
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7

Apoptosis Assay in Tumor Tissue

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Tumor tissue apoptosis was investigated with the TUNEL assay (Beyotime, Jiangsu, China), according to the manufacturer’s instructions. The stained tissues were visualized under a fluorescence microscope.
Ma B., Zhang H., Wang Y., Zhao A., Zhu Z., Bao X., Sun Y., Li L, & Zhang Q. (2018). Corosolic acid, a natural triterpenoid, induces ER stress-dependent apoptosis in human castration resistant prostate cancer cells via activation of IRE-1/JNK, PERK/CHOP and TRIB3. Journal of Experimental & Clinical Cancer Research : CR, 37, 210.
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8

Comprehensive Analysis of Cell Proliferation and Apoptosis

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Cell proliferation was analyzed using a Cell-Light 5-ethynyl-2′-deoxyuridine (EdU) Apollo567 In Vitro Kit (RiboBio, Shanghai, China) in strict accordance with the manufacturer's instructions. Images were acquired at 20 × magnification with a fluorescence microscope (Leica, Solms, Germany). EdU-positive cells were counted in the selected cells and calculated as the mean value from multiple fields of view. Cell apoptosis assays were performed using an Annexin V-PE 7-AAD Apoptosis Detection Kit (BD Biosciences, CA, USA) and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL assay, Beyotime, Shanghai, China) according to the manufacturer's protocol. For Annexin V-PE 7-AAD Apoptosis assay, the cells were analyzed the percentage of apoptotic cells, that cells undergoing early and late apoptosis, among 10,000–20,000 counted cells using a flow cytometer (BD Biosciences, CA, USA). For TUNEL assay, images of apoptosis cells were obtained using a microscope (Leica, Solms, Germany).
Wang X.F., Li M.L., Fang Q.Q., Zhao W.Y., Lou D., Hu Y.Y., Chen J., Wang X.Z, & Tan W.Q. (2020). Flexible electrical stimulation device with Chitosan-Vaseline® dressing accelerates wound healing in diabetes. Bioactive Materials, 6(1), 230-243.
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9

Apoptosis Detected by TUNEL Assay

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Apoptosis was detected by using TUNEL assay (Beyotime Biotechnology, Shanghai, China). The nuclei were stained again with 4,6-diamino-2-phenylindole (DAPI; Beyotime Biotechnology, Shanghai, China). Three random regions of each group of cells were observed under a fluorescence microscope (Leica, DE).
Zheng Z., Yu Z., Xu B., Zhou Y., Xing Y., Li Q., Tang W, & Peng F. (2022). Pretreatment with Shenmai Injection Protects against Coronary Microvascular Dysfunction. Evidence-based Complementary and Alternative Medicine : eCAM, 2022, 8630480.
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10

Apoptosis Detection via TUNEL Assay

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Fixed cells were used for TUNEL assay (Beyotime, Shanghai, China). Briefly, the cells were incubated with 50 μl of TUNEL reaction buffer for 50 min and diaminobenzidine (DAB) for 3 min, the results were photographed using an epifluorescence microscope at ×400 magnification (Nikon Eclipse 80i).
Zhou J., Wang Y., Zhang L., Chen Q., Zhu X., Jiang P., Jiang N., Zhao W, & Li B. (2022). Engineered Exosomes-Mediated Transfer of hsa-miR-320a Overcomes Chemoresistance in Cervical Cancer Cells via Targeting MCL1. Frontiers in Pharmacology, 13, 883445.
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