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Tunel assay

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The TUNEL (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) assay is a method used to detect and quantify apoptosis, or programmed cell death, in cells. The assay works by labeling the fragmented DNA in apoptotic cells, allowing for their identification and analysis.

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Close spelling variants of this product

In situ TUNEL assay kit Diagnosis TUNEL assay kit Terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) assay kit One-step TUNEL assay kit One-step TUNEL Apoptosis Assay KIT Tunel assay kit for apoptosis TUNEL assay kit TUNEL apoptosis assay TdT-mediated dUTP nick end labeling (TUNEL) apoptosis assay kit TUNEL fluorescein assay kit

326 protocols using tunel assay

1

Apoptosis Analysis in Cardiac Cells

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Apoptosis was analyzed by TUNEL assay (Roche) and Hoechst 33258 staining (H1399; Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. The TUNEL assay was used in order to detect DNA strand breaks. Briefly, the NRCs were grown on laminin-coated chamber slides and exposed to LPS challenge as described above. Twenty-four hours later, the cells were fixed in 4% paraformaldehyde (PFA) for 1 h, and the TUNEL assay was performed, according to the manufacturer's instructions using a commercial kit (Roche). Apoptotic nuclei stained with fluorescein isothiocyanate (FITC; TUNEL assay) were visualized by confocal microscopy (FV1000; Olympus, Tokyo, Japan). TO-PRO®-3 stain was used to stain all nuclei. TUNEL-positive cells were counted and the number of apoptotic cells was expressed as a percentage of the total number of cells.
Zhang X., Zhang X., Xiong Y., Xu C., Liu X., Lin J., Mu G., Xu S, & Liu W. (2016). Sarcolemmal ATP-sensitive potassium channel protects cardiac myocytes against lipopolysaccharide-induced apoptosis. International Journal of Molecular Medicine, 38(3), 758-766.
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2

Apoptosis Quantification in Tumor Tissue

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In situ tumor cell apoptosis was determined using the TdT-mediated dUTP nick-end labeling (TUNEL) assay (Boehringer-Mannheim). Sectioned tumor tissue was embedded in paraffin. Three slides from each group were evaluated for the apoptotic cells. Six slide fields were randomly examined using a defined rectangular field area with × 200 magnification, and apoptotic cells were counted in each field.
Zhang M., Qian J., Lan Y., Lu Y., Li H., Hong B., Zheng Y., He J., Yang J, & Yi Q. (2014). Anti-β2M monoclonal antibodies kill myeloma cells via cell- and complement-mediated cytotoxicity. International journal of cancer. Journal international du cancer, 135(5), 1132-1141.
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3

Evaluating PEDOT/Zn Micromotor Toxicity

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To evaluate the toxicity of PEDOT/Zn micromotors in vivo, ICR male mice at 6 weeks of age were orally administered with 0.3 mL of the PEDOT/Zn micromotors, as well as with free AuNPs or AuNPs-loaded PEDOT/Zn micromotors. Mice administered with PBS were used as a negative control. At 6 h after the administration, the mice were sacrificed and the stomachs were collected for histological analysis. The longitudinal sections of the gastric tissue were fixed in neutral-buffered 10% formalin and then embedded in paraffin. The tissue sections were stained with hematoxylin and eosin (H&E). Epithelial cell apoptosis was evaluated by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) assay (Boehringer Mannheim, Indianapolis, IN). Sections were visualized by the Hamamatsu NanoZoomer 2.0HT, and the images were processed using NDP viewing software. All animal experiments were in compliance with institutional animal use and care regulations.
Gao W., Dong R., Thamphiwatana S., Li J., Gao W., Zhang L, & Wang J. (2014). Artificial Micromotors in the Mouse’s Stomach: A Step toward in Vivo Use of Synthetic Motors. ACS Nano, 9(1), 117-123.
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4

Evaluation of NAFLD Liver Pathology

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Formalin-fixed, paraffin-embedded liver biopsies were cut into 5 μm serial sections. NAFLD severity was assessed with hematoxylin and eosin staining using criteria described by Brunt et al. [22 (link)].
For immunohistochemistry, sections were deparafinized with xylene and rehydrated. Slides were incubated in 3% hydrogen peroxide/methanol. Antigen retrieval was performed by heating in 10 mM sodium citrate (pH 6.0). Sections were incubated overnight with caspase-2 antibody. Horseradish peroxidase-conjugated IgG secondary antibody was used. Antigens were demonstrated by diaminobenzidine (K3466; Dako Envision, Carpinteria, CA). Omitting primary antibodies from reactions eliminated staining, demonstrating specificity. Tissue sections were counterstained with Aqua Hematoxylin-INNOVEX (Innovex Biosciences). Morphometric analysis was done with Metamorph Software (Molecular Devices Corporation, Downingtown, PA). Immunohistochemistry antibodies are specified in Supplemental Table 1.
Liver fibrosis was assessed by Picrosirius red (Sigma, ST. Louis, MO) staining. Quantification was done by morphometric analysis in randomly chosen sections (x10 magnification, 20 fields/sample) [23 (link)].
Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay (Boehringer Mannheim, Germany) was performed according to the manufacturer's suggestions.
Machado M., Michelotti G., Pereira de Almeida T., Boursier J., Kruger L., Swiderska-Syn M., Karaca G., Xie G., Guy C., Bohnic B., Lindblom K., Johnson E., Kornbluth S, & Diehl A. (2014). Reduced Lipoapoptosis, Hedgehog Pathway Activation, and Fibrosis in Caspase-2 deficient Mice with Nonalcoholic Steatohepatitis. Gut, 64(7), 1148-1157.
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5

Evaluating Skin Toxicity of AuC-Liposome Hydrogel

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To evaluate the skin toxicity of AuC–liposome hydrogel, ICR mice at 6 weeks of age were purchased from Charles River Laboratories. Mice were shaved on the back 24 h prior to the study. Then the AuC–liposome hydrogel was applied on the shaved area once daily for a period of 7 days. Mice treated with PBS served as a negative control. To prevent the gel from drying out, the skin was covered with gauzes. Twenty-four hours after the last topical administration, the mice were sacrificed and the skin was cross-sectioned by 8 mm biopsy punch for histological examination. The skin tissues from each mouse were fixed in buffered 10% formalin for 18 h and then embedded in paraffin. The tissue sections were stained with hematoxylin and eosin (H&E). Epithelial cell apoptosis was evaluated by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) assay (Boehringer Mannheim, Indianapolis, IN). Sections were visualized by Hamamatsu Nanozommer 2.0HT. Images were processed using NDP viewer software. Five mice were used for each test group (n = 5).
Gao W., Vecchio D., Li J., Zhu J., Zhang Q., Fu V., Li J., Thamphiwatana S., Lu D, & Zhang L. (2014). Hydrogel Containing Nanoparticle-Stabilized Liposomes for Topical Antimicrobial Delivery. ACS Nano, 8(3), 2900-2907.
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6

Cardiac Myocyte Apoptosis Quantification

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In situ cardiac myocyte apoptosis was examined by the TdTmediated dUTP nick end labelling (TUNEL) assay (Boehringer Mannheim, Indianapolis, IN, USA) as described previously [12] . Five slides from each block were evaluated for percentage of apoptotic cells by using the TUNEL assay. Then 5 watch fields were chosen randomly under microscope on each section. Positive brown cells and total cells were counted by MIAS4.0 (Medical Image Analysis System, Beijing Bingyang Keji Corp., China). The 5 fields were averaged for each block. Then the blocks were averaged for each heart to obtain one number for each heart. Apoptosis index (positive cells/total cells×100%) was used as the indicator of apoptosis.
Hu X., Ma R., Lu J., Zhang K., Xu W., Jiang H, & Da Y. (2016). IL-23 Promotes Myocardial I/R Injury by Increasing the Inflammatory Responses and Oxidative Stress Reactions. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 38(6).
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7

Apoptosis Detection in Brain Tissue

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Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) detection is widely used for apoptosis. Following the manufacturer's instructions, brain tissue was stained with TUNEL assay (Roche, Basel, Switzerland) for an hour and then examined under a fluorescence microscope (Olympus, Japan).
Cong P., Tong C., Liu Y., Shi L., Shi X., Zhao Y., Xiao K., Jin H., Liu Y, & Hou M. (2019). CD28 Deficiency Ameliorates Thoracic Blast Exposure-Induced Oxidative Stress and Apoptosis in the Brain through the PI3K/Nrf2/Keap1 Signaling Pathway. Oxidative Medicine and Cellular Longevity, 2019, 8460290.
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8

Evaluating Tumor Angiogenesis and Apoptosis

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Expression of CD31 in tumors was assessed by IHC staining as described previously.22 In brief, paraffin‐embedded tumor sections were incubated with anti‐CD31 antibody (1:100) overnight at 4°C, and then incubated with secondary antibody.
Apoptosis was evaluated by TUNEL assay (Roche diagnostics, Mannheim, Germany) following the manufacturer's instructions. The image of each case was captured using a fluorescence microscope (Nikon Eclipse‐Ti; Nikon, Tokyo, Japan) and analyzed using Image‐Pro Plus 6.0 (IPP6) software.
Li X., Chen C., Dai Y., Huang C., Han Q., Jing L., Ma Y., Xu Y., Liu Y., Zhao L., Wang J., Sun X, & Yao X. (2019). Cinobufagin suppresses colorectal cancer angiogenesis by disrupting the endothelial mammalian target of rapamycin/hypoxia‐inducible factor 1α axis. Cancer Science, 110(5), 1724-1734.
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9

Apoptosis Assessment of Mouse Keratinocytes and Human PV Biopsies

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Mouse keratinocytes and biopsies from 8-week-old C57BL/6J mice and human PV patients [6 (link),23 (link)] were subjected to a TUNEL assay (Roche Diagnostics, Basel, Switzerland) according to the manufacturer instructions. FACS analyses were performed as described [6 (link)].
Luyet C., Schulze K., Sayar B.S., Howald D., Müller E.J, & Galichet A. (2015). Preclinical Studies Identify Non-Apoptotic Low-Level Caspase-3 as Therapeutic Target in Pemphigus Vulgaris. PLoS ONE, 10(3), e0119809.
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10

Infection Induced Cellular Response

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THP-1 cells (TIB-202, ATCC) were treated with 8 ng/ml phorbol myristate acetate (Sigma-Aldrich) to differentiate into macrophage-like cells. THP-1 or A549 human lung adenocarcinoma epithelial cell line (CCL-185, ATCC) cells were infected with a multiplicity of infection (MOI) of 10:1 (bacteria:mammalian cells). Apoptosis of THP-1 cells (using TUNEL assay, Roche) and necrosis of A549 cells (using lactose dehydrogenase release assay, Roche) were determined at 96 hours post infection.
Smith K.L., Saini D., Bardarov S., Larsen M., Frothingham R., Gandhi N.R., Jacobs Jr. W.R., Sturm A.W, & Lee S. (2014). Reduced Virulence of an Extensively Drug-Resistant Outbreak Strain of Mycobacterium tuberculosis in a Murine Model. PLoS ONE, 9(4), e94953.
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