Click it plus tunel assay

Manufactured by Thermo Fisher Scientific

Sourced in United States, France, Germany, United Kingdom

The Click-iT Plus TUNEL Assay is a fluorescence-based method for detecting DNA fragmentation, a hallmark of apoptosis. The assay utilizes a modified nucleotide, EdUTP, which is incorporated into the DNA breaks. The incorporated EdUTP is then detected using a fluorescent azide dye, providing a quantitative measure of DNA fragmentation.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

TUNEL assay PLUSTM

111 protocols using click it plus tunel assay

In Situ Apoptosis Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols

Slides were dried for 1 h before staining with Click-iT Plus TUNEL assay for in situ apoptosis detection (Fisher; C10618). Steps followed the manufacturer's instructions. A positive control slide was treated with DNase I.

Rosko L.M., Gentile T., Smith V.N., Manavi Z., Melchor G.S., Hu J., Shults N.V., Albanese C., Lee Y., Rodriguez O, & Huang J.K. (2023). Cerebral Creatine Deficiency Affects the Timing of Oligodendrocyte Myelination. The Journal of Neuroscience, 43(7), 1143-1153.

+ Open protocol

+ Expand

Colon and Stomach Cancer Cell Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

The human colon cancer cell line (CaCo2) and stomach cancer cell line (AGS) were purchased from ATCC (Manassas, Virginia). 5FU, sodium dodecyl sulfate, and Coumarin 6 were purchased from Sigma-Aldrich (St. Luis, Missouri). β-Glucan (MW 179,000) was purchased from Megazyme (Wicklow, Ireland). Bcl2 siRNA was purchased from Cell Signaling Technology (Danvers, Massachusetts). Fetal bovine serum (FBS), penicillin, phosphate-buffered saline (PBS), and 0.05% Trypsin–EDTA were purchased from Life Technologies (Carlsbad, CA). Bcl-2 SiRNA I (Cat #6441) was purchased from Cell Signaling Technology (Danvers, MA). Bcl2 antibody, Ki-67 antibody, and FITC-Goat anti-Rabbit IgG were purchased from ABclonal (Woburn, Massachusetts). Simulated gastric juice was purchased from RICCA (Arlington, Texas). Click-iT Plus TUNEL assay, eosin, xylene, Harris hematoxylin, was purchased from Fisher Scientific (Waltham, Massachusetts).

Loparev V.N., McCaustland K., Holloway B.P., Krause P.R., Takayama M, & Schmid D.S. (2000). Rapid Genotyping of Varicella-Zoster Virus Vaccine and Wild-Type Strains with Fluorophore-Labeled Hybridization Probes. Journal of Clinical Microbiology, 38(12), 4315-4319.

+ Open protocol

+ Expand

TUNEL Assay for Tissue Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols

TUNEL assay was performed using the Click-iT Plus TUNEL assay (Molecular Probes; cat. no. C10619) following guidelines for cells with the following differences: permeabilization of tissues was performed overnight in PBS+0.3% Triton X-100, TdT reaction was performed by incubating overnight followed by 1.5 h at 37 °C, and incubation with Click-iT reagent was performed for 45 min at 37 °C. TUNEL experiments were performed two times.

Manent J., Banerjee S., de Matos Simoes R., Zoranovic T., Mitsiades C., Penninger J.M., Simpson K.J., Humbert P.O, & Richardson H.E. (2017). Autophagy suppresses Ras-driven epithelial tumourigenesis by limiting the accumulation of reactive oxygen species. Oncogene, 36(40), 5576-5592.

+ Open protocol

+ Expand

Quantifying Brain Cell Apoptosis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell apoptosis was measured by quantifying TUNEL⁺ cells (Terminal deoxynucleotidyl transferase dUTP nick end labeling) in brain sections using a commercial kit (Click-iT^™ Plus TUNEL assay, Molecular Probes). TUNEL⁺ cells were detected at an emission of 647 nm and quantified as the ratio to DAPI-positive nuclei, neuronal marker NeuN, or oligodendrocyte marker O4 in at least five randomly selected visual fields for a group. The quantification of the double-positive TUNEL cells was analyzed by MetaMorph software (Molecular Devices).

Kuan C.Y., Chen H.R., Gao N., Kuo Y.M., Chen C.W., Yang D., Kinkaid M.M., Hu E, & Sun Y.Y. (2020). Brain-Targeted Hypoxia-Inducible Factor Stabilization Reduces Neonatal Hypoxic-Ischemic Brain Injury. Neurobiology of disease, 148, 105200.

+ Open protocol

+ Expand

Apoptosis and Autophagy in Gonads

Check if the same lab product or an alternative is used in the 5 most similar protocols

In paraffin sections, apoptosis was assessed by TUNEL technique, using Click-iT Plus TUNEL Assay for in situ apoptosis detection with Alexa Fluor 647 dye (Molecular Probes, Life Technologies). The procedure was performed according to manufacturer’s instructions. In four gonads, TUNEL assay was followed by immunofluorescence staining for LC3 according to the procedure described above. Each gonad was evaluated in several sections for presence of apoptotic cells, autophagy and micronuclei in gonocytes. Documentation of results was done using Olympus FV1000 confocal microscope.

Chmielewska M., Dedukh D., Haczkiewicz K., Rozenblut-Kościsty B., Kaźmierczak M., Kolenda K., Serwa E., Pietras-Lebioda A., Krasikova A, & Ogielska M. (2018). The programmed DNA elimination and formation of micronuclei in germ line cells of the natural hybridogenetic water frog Pelophylax esculentus. Scientific Reports, 8, 7870.

+ Open protocol

+ Expand

Caspase-3/7 Activity Determination in HepaRG Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The activity of caspase-3/7 (CPP32/apopain)-like proteases was determined using the EnzChek Caspase-3 Assay Kit following manufacturer instructions. Briefly, 5 × 10 5 HepaRG cells on 6-well plates were incubated with or without LAs following the same procedures as the MTT assay. After 48 hours, the cells were washed with PBS, lysed, and caspase activity in the extracts was measured. Fluorescent product of the Z-DEVDrhodamine 110 substrate generated by caspase-3-like proteases was detected by a Polarstar Omega fluorometer with excitation/emission at 496/520 nm. Background fluorescence was determined by following the same procedures without cells and subtracted from the total. Negative control was performed by including a specific caspase-3 inhibitor (Ac-DEVD-CHO) in HepaRG cells. Positive control was performed by treating HepaRG cells with doxorubicin (50 ng/mL). 18 Apoptosis was also detected in situ by using a Click-iT Plus TUNEL assay (Molecular Probes, Villebon-sur-Yvette, France). Results were expressed in percentage of marked cell by dividing the number of stained cells by the number of nuclei assessed by Hoechst.

Le Gac G., Angenard G., Clément B., Laviolle B., Coulouarn C, & Beloeil H. (2017). Local Anesthetics Inhibit the Growth of Human Hepatocellular Carcinoma Cells. Anesthesia and analgesia, 125(5).

+ Open protocol

+ Expand

Apoptosis Analysis by TUNEL Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Apoptosis was analyzed by TUNEL assay using Click-iT^® Plus TUNEL Assay (Life Technologies, Inc., Carlsbad, CA, USA) according to manufacturer’s instruction. Briefly, at the end of the indicated treatments, primary rat cerebral cortical neurons grown on coverslips were incubated with TdT reaction mixture for 2 h at 37°C, followed by 30-min incubation with the Alexa Fluor^® 594 dye. Then, the cells were counterstained with DAPI (Sigma-Aldrich) for 20 min and observed under a fluorescence microscope (Leica, Germany). The TUNEL-positive nuclei of six non-overlapping fields per coverslip were counted by a researcher blinded to treatment, and these counts were converted to percentages by comparing the TUNEL-positive counts to the total number of cell nuclei as determined by DAPI counterstaining, that is TUNEL-positive ratio = (number of red nuclei/number of blue nuclei) × 100%.

Zhang Y., Yang K., Wang T., Li W., Jin X, & Liu W. (2017). Nrdp1 Increases Ischemia Induced Primary Rat Cerebral Cortical Neurons and Pheochromocytoma Cells Apoptosis Via Downregulation of HIF-1α Protein. Frontiers in Cellular Neuroscience, 11, 293.

+ Open protocol

+ Expand

TUNEL Assay for Apoptosis Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols

The TUNEL assay was conducted utilizing the Click-iT^® Plus TUNEL Assay (Life technologies, USA) according to the manufacturer’s protocol. Briefly, HEI-OC1 cells that were given the indicated treatment were fixed with 4% PFA in PBS for 15 min at room tempreture and then washed with PBS. Then, the cells were permeabilized with 0.1% Triton X-100 in PBS for 10 min, and stained with TUNEL working solution for 1 h at 37 °C in dark. Cell nuclei were stained with DAPI. The specimens were visualized using a Leica confocal laser scanning microscopy.

Yin H., Sun G., Yang Q., Chen C., Qi Q., Wang H, & Li J. (2017). NLRX1 accelerates cisplatin-induced ototoxity in HEI-OC1 cells via promoting generation of ROS and activation of JNK signaling pathway. Scientific Reports, 7, 44311.

+ Open protocol

+ Expand

Apoptosis Identification Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Apoptotic cells were identified for appearance of apoptotic morphology including membrane blebbing and cell rounding. Live cells retained their normal spread morphology. Apoptotic cells were also determined by anti-Annexin V immunostaining. The Life Technologies™ Click-iT® Plus TUNEL assay with Alexa Fluor® 647 dye was used to detect the fragmented DNA.

Qin R., Melamed S., Yang B., Saxena M., Sheetz M.P, & Wolfenson H. (2022). Tumor Suppressor DAPK1 Catalyzes Adhesion Assembly on Rigid but Anoikis on Soft Matrices. Frontiers in Cell and Developmental Biology, 10, 959521.

+ Open protocol

+ Expand

TUNEL Assay for Apoptosis Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols

TUNEL reactions were performed on paraffin sections from 2 dpf embryos. Slides were deparaffinized, treated with sodium citrate solution as above, and labeling was performed using the Click-iT Plus TUNEL Assay (Life Technologies Cat. no. C10617), according to manufacturer instructions. Samples were permeabilized with proteinase K for 30 min at 37°C. A positive control was generated by treatment with a 1:50 dilution of DNAseI (ThermoFisher Scientific Cat. no. EN0525) in reaction buffer, followed by incubation at room temperature for 30 min. TUNEL assays were performed on experimental and positive control slides simultaneously, then slides were mounted with VectaShield-DAPI media (Vector Laboratories, H-1400).

Timoshevskiy V.A., Herdy J.R., Keinath M.C, & Smith J.J. (2016). Cellular and Molecular Features of Developmentally Programmed Genome Rearrangement in a Vertebrate (Sea Lamprey: Petromyzon marinus). PLoS Genetics, 12(6), e1006103.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!