For TUNEL assays, ovaries were collected from Hdac3flox/flox and Hdac3cKO mice and fixed in cold 4% PFA for 1 h. The tissues were embedded in paraffin wax and serially sectioned at 5 μm. Apoptotic cells were detected using a TUNEL Apoptosis Detection Kit (Invitrogen, Life Technologies, USA) after HDAC3 immunostaining.
Tunel apoptosis detection kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The TUNEL Apoptosis Detection kit is a laboratory tool used to detect and quantify apoptosis, a type of programmed cell death, in samples. The kit utilizes the TUNEL (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) assay, which labels DNA strand breaks that occur during apoptosis, allowing for their identification and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescence microscope, by Olympus (2 mentions)
Optical microscope, by Olympus (1 mentions)
Hematoxylin, by Solarbio (1 mentions)
Luteolin, by Abcam (1 mentions)
Cell culture reagents, by Merck Group (1 mentions)
Jc 1 dye, by Thermo Fisher Scientific (1 mentions)
Caspase 3 assay kit, by Merck Group (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Polybrene, by Merck Group (1 mentions)
Puromycin, by Merck Group (1 mentions)
5 protocols using tunel apoptosis detection kit
1
HDAC3 Regulation of Ovarian Apoptosis
2
Evaluating Apoptosis in Transfected SKOV3 Cells
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The transfected SKOV3 cells were fixed with 4% paraformaldehyde at 4˚C for 20 min. Cell apoptosis was examined with a TUNEL Apoptosis Detection kit (Invitrogen; Thermo Fisher Scientific, Inc.) at 37˚C for 1 h, according to the manufacturer's instructions. Subsequently, cells were incubated with diaminobenzene for 10 min and counterstained with hematoxylin (Beijing Solarbio Science & Technology Co., Ltd.) for 30 sec at room temperature. The apoptosis of TUNEL-positive cells was observed under an optical microscope (magnification, x200; Olympus Corporation) and cells were counted in five randomly selected microscopic fields.
3
Evaluating Apoptosis with TUNEL Assay
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For the evaluation of apoptosis, a TUNEL Apoptosis Detection kit (Invitrogen, Carlsbad, Calif, USA) was employed in this study following manufacturer’s recommendations. Raw 264.7 cells were fixed with 4% paraformaldehyde and permeabilized by 0.3% Triton X-100. Cells were then incubated with 50 μl TUNEL reaction buffer. The nuclei were counterstained with DAPI in the dark. Images were visualized and captured using a fluorescence microscope (Olympus Corporation).
4
Luteolin-Induced Apoptosis and ROS
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Luteolin (purity > 98%, ab-120662, Figure 1(a) ) was obtained from Abcam (Cambridge, UK). All antibodies were provided by Cell Signaling Technology (Shanghai, China) and Santa Cruz Biotech (Santa Cruz, CA). Carboxy-2′,7′-dichlorodihydrofluorescein diacetate (carboxy-H2DCFDA) fluorescent dye, TUNEL apoptosis detection kit (FITC-labelled), JC-1 dye, TRIzol reagent, and Lipofectamine 2000 were obtained from Invitrogen (Shanghai, China) and ThermoFisher Scientific (San Jose, CA). Puromycin, polybrene, the Annexin V-fluorescent-activated cell sorting (FACS) assay kit, the caspase-3 assay kit, and cell culture reagents were provided by Sigma-Aldrich (St. Louis, MO, USA).
5
Evaluating Apoptosis via TUNEL Assay
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For the evaluation of apoptosis, a TUNEL Apoptosis Detection kit (Invitrogen, Carlsbad, CA, USA) was employed in this study following manufacturer's recommendations. SW1353 cells were fixed with 4% paraformaldehyde and then incubated with 0.1% Triton X-100 to permeabilize the cell membrane. Cells were then incubated with 50 μL TUNEL reaction buffer for 1 h at 37 °C. The nuclei were counterstained with DAPI in the dark and the slides were then mounted with anti-fade mounting medium. Images were visualized and captured using a fluorescence microscope (Olympus Corporation).
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