MPC5 cell apoptosis was detected by TUNEL assay using the TUNEL Apoptosis Assay Kit (Beyotime, Haimen, China). The cells were incubated in terminal deoxynucleotidyl transferase (TDT)-mediated dUTP-biotin nick end-labeling (TUNEL). The TUNEL positive nuclei were observed by optical microscope (Leica DM4P, Shanghai, China).
Tunel apoptosis assay kit
Manufactured by Beyotime
Sourced in China, Japan
The TUNEL Apoptosis Assay Kit is a laboratory reagent used to detect and quantify apoptosis, a type of programmed cell death, in cell samples. The kit utilizes the TUNEL (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) technique to label DNA strand breaks, a hallmark of apoptosis. It provides a reliable method for the assessment of apoptosis in various cell types and experimental settings.
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Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Beyotime (6 mentions)
Fluorescence microscope, by Olympus (6 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (5 mentions)
Annexin 5 fitc apoptosis detection kit, by Beyotime (4 mentions)
Cell counting kit 8 cck 8, by Beyotime (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Cleaved caspase 3, by Cell Signaling Technology (3 mentions)
Antifade mounting medium with dapi, by Beyotime (3 mentions)
Bcl 2, by Cell Signaling Technology (3 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (3 mentions)
Triton x 100, by Beyotime (3 mentions)
Cell counting kit 8 (cck8), by Sangon (2 mentions)
Fluorescence microscope, by Nikon (2 mentions)
Dnase free proteinase k, by Beyotime (2 mentions)
Hoechst 33258, by Beyotime (2 mentions)
Dapi staining kit, by Beyotime (2 mentions)
β actin antibody, by Proteintech (2 mentions)
Antifade mounting medium, by Beyotime (2 mentions)
C1091, by Beyotime (2 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (2 mentions)
193 protocols using tunel apoptosis assay kit
1
Detecting MPC5 Cell Apoptosis by TUNEL
2
Tumor Inhibition and Apoptosis Analysis
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After the last administration, the mice were fasted for 12 h, and all of the experimental mice were weighed. The tumor was carefully removed and weighed immediately. The tumor inhibition rate was calculated as follows:
Note: M1, the average tumor weights of the tumor model group; M2, the average tumor weights of the treated group.
Tumor tissue was fixed with 4% paraformaldehyde, then dehydrated, embedded, sectioned and stained with HE. The status of tissue necrosis was observed under a microscope(Eclipse TS100; Nikon Corporation, Tokyo, Japan).TUNEL (TdT-mediated dUTP Nick-End Labeling) assay was performed to detect apoptosis of tumor cells, using the TUNEL Apoptosis Assay Kit (Beyotime Institute of Biotechnology) according to the manufacturer’s instructions. The results was observed under a fluorescence microscope (TI, Nikon Corporation, Tokyo, Japan) at × 200 magnification.
Note: M1, the average tumor weights of the tumor model group; M2, the average tumor weights of the treated group.
Tumor tissue was fixed with 4% paraformaldehyde, then dehydrated, embedded, sectioned and stained with HE. The status of tissue necrosis was observed under a microscope(Eclipse TS100; Nikon Corporation, Tokyo, Japan).TUNEL (TdT-mediated dUTP Nick-End Labeling) assay was performed to detect apoptosis of tumor cells, using the TUNEL Apoptosis Assay Kit (Beyotime Institute of Biotechnology) according to the manufacturer’s instructions. The results was observed under a fluorescence microscope (TI, Nikon Corporation, Tokyo, Japan) at × 200 magnification.
3
Investigating S1P Signaling in Liver Fibrosis
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Sal was procured from Shanghai Tauto Biotechnology (Shanghai, China), the alanine transaminase (ALT), aspartate aminotransferase (AST), and hydroxyproline (Hyp) detection kits were acquired from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). FN, type I collagen (Col I), Bax and Bcl-2 antibodies were obtained from Abcam (Cambridge, United Kingdom). JNK, p-JNK, PARP, and cleaved caspase 3 antibodies were purchased from Cell Signaling Technology (Massachusetts, United States). SphK1, SphK2, S1PR1, S1PR2, S1PR3, CD9, and tumor susceptibility gene 101 (TSG101) antibodies were procured from ImmunoWay Biotechnology Company (Suzhou, China). Antibody against GAPDH was provided by Proteintech (Wuhan, China). TdT-mediated dUTP nick end labeling (TUNEL) Apoptosis Assay Kit, 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DPAI), and Cy3-labeled goat anti-rabbit IgG were all bought from Beyotime Institute of Biotechnology (Shanghai, China). The S1P Assay Kit (S1P-ELISA) was delivered by Echelon Biosciences (Salt Lake City, United States).
4
Cell Proliferation and Apoptosis Assays
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The Cell Counting Kit-8 (Sangon, China) and TUNEL Apoptosis Assay Kit (Beyotime, China) were applied to perform cell proliferation and apoptosis assessments. The instructions for kits were strictly followed.
5
Apoptosis Assessment in Tumor Samples
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The tumor samples were fixed with 4% paraformaldehyde, embedded in paraffin, and sliced into 4-μm sections. A TUNEL Apoptosis Assay Kit (Beyotime, China) was applied for apoptosis rate analyses following the manufacturer’s instructions. For IHC staining, the sections were incubated with primary and secondary antibodies consecutively. After DAB staining, the sections were observed under a microscope (Olympus, Japan). The IHC score was calculated according to a previous study (23 (link)).
6
TUNEL Assay for Apoptosis Detection
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A One Step dUTP nick-end labeling (TUNEL) Apoptosis Assay Kit (Beyotime, Shanghai, China) was used to detect cell apoptosis according to the manufacturer’s instructions. Briefly, the cells were incubated with 50 μl TUNEL detection mixture at 37°C for 1 h in the dark and rinsed in PBS. Subsequently, 50 μl streptavidin–horseradish peroxidase (HRP) solution was added and incubated for 30 min at room temperature. DAB staining was then performed, followed by hematoxylin staining of the nuclei.
7
Apoptosis Assays for Fish Erythrocytes
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The PS exposure and DNA fragmentation in fish erythrocytes were respectively assessed by the Annexin V-FITC Apoptosis Detection Kit and TdT-mediated dUTP nick end labeling (TUNEL) Apoptosis Assay Kit (Beyotime, Nantong, China) as described previously (Li et al., 2016 (link)).
8
Multifunctional Nanomaterial Synthesis and Evaluation
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CTAC, TEA, xanthine, and xanthine oxidase (XO) were purchased from Sigma-Aldrich. TEOS and NaOH were obtained from Shanghai Lingfeng Chemical Reagent Co., Ltd. Fe(acac)3 was provided by J&K Scientific. Urea, RhB, iron chloride hexahydrate (FeCl3·6H2O) and 3,3′,5,5′-tetramethyl-benzidine (TMB) were bought from Sinopharm Chemical Reagents Co., Ltd. Methoxy PEG silane was purchased from Shanghai Yare Biotech, Co., Ltd. Gallic acid was acquired from Macklin. Dulbecco’s modified eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin/streptomycin, and PBS were provided by Gibco. FITC, 4′, 6-diamidino-2-phenylindole (DAPI), DCFH-DA, CCK-8 assay, calcein-AM, PI, annexin V-FITC, H&E, paraformaldehyde (PFA) and TUNEL apoptosis assay kit were acquired from Beyotime. Primary antibody against GPX4 and a biotinylated secondary antibody (anti-rabbit IgG, HRP-linked antibody) were purchased from Cell Signaling Technology.
9
TUNEL Assay for Apoptosis Detection
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All operations were performed according to the instructions provided with the TUNEL Apoptosis Assay kit (Beyotime Institute of Biotechnology). After returning the frozen brain tissue sections to room temperature, they were fixed in 4% paraformaldehyde for 30 min at room temperature, and permeabilized with PBS containing 0.5% Triton X-100 for 5 min. The tissues were then incubated with TUNEL reaction mixture for 60 min at 37°C, protected from light. After blocking with mounting medium containing 4,6-diamidino-2-phenylindole (DAPI; Beijing Solarbio Science & Technology Co., Ltd.), five random fields of view were observed under a fluorescence microscope (Nikon Corporation; magnification, ×200).
10
TUNEL Assay for Apoptosis Detection
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TUNEL Apoptosis Assay Kit (C1086, provided by Beyotime, China) was utilized in this research. The paraffin sections were subjected to treatment with dewaxing and rehydration followed by a reaction with DNase-free proteinase K (ST532, Beyotime, China) at 37°C for 20 min. After washing thoroughly, the sections were exposed to the prepared TUNEL detection solution at 37°C away from light for 1 h. After washing, the sections were blocked with Antifade Mounting Medium with DAPI (P0131, Beyotime, China) and then observed in the fluorescence microscopy with its excitation and emission wavelength at 500 and 565 nm, respectively.
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