TUNEL assays were performed on the UT-SCC-60A and UT-SCC-60B cells using an In Situ Cell Death Detection Kit with Fluorescein (Roche, Basel, Switzerland) according to the manufacturer’s instructions. The fixed cells were treated with DNase I as positive controls. Samples were incubated with the TUNEL reaction mixture for 1 hr at 37°C in the dark and then washed twice in PBS. The condensed or fragmented nuclei of apoptotic cells were observed under an FV3000 Confocal Laser Scanning Microscope.
In situ cell death detection kit with fluorescein
Manufactured by Roche
Sourced in Germany, United States
The In Situ Cell Death Detection Kit with Fluorescein is a laboratory product designed to detect and quantify cell death in a variety of samples. The kit utilizes the TUNEL (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) method to label DNA strand breaks, a hallmark of apoptosis, or programmed cell death. The kit provides a fluorescent labeling system that can be used to visualize and analyze cell death in a range of biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 780 confocal microscope, by Zeiss (3 mentions)
Dako envision kit hrp, by Agilent Technologies (2 mentions)
Lsm 780, by Zeiss (2 mentions)
Cryostat microtome, by Leica (1 mentions)
Anti rabbit igg fitc, by Merck Group (1 mentions)
Anti mouse igg tritc, by Merck Group (1 mentions)
Cryomicrotome, by Leica (1 mentions)
Celltiter 96 aqueous one solution cell kit, by Promega (1 mentions)
Celltiter glo luminescent cell viability assay, by Promega (1 mentions)
Permount, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Human tubal fluid, by Irvine Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Vectashield antifade mounting medium with dapi, by Vector Laboratories (1 mentions)
Anti cleaved caspase 3 primary antibody, by Cell Signaling Technology (1 mentions)
Alexa fluor 594, by Thermo Fisher Scientific (1 mentions)
Inverted light microscope, by Zeiss (1 mentions)
Sp5 confocal microscope, by Leica (1 mentions)
18 protocols using in situ cell death detection kit with fluorescein
1
Detecting Apoptosis in Cancer Cells
2
Quantifying Neuronal Apoptosis Post-ICH
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To quantify neuronal apoptosis events, double staining with terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and the neuron marker NeuN was conducted with an In Situ Cell Death Detection Kit with Fluorescein (Roche, Germany) at 72 h after ICH according to the manufacturer's instructions. The number of TUNEL-positive neurons was counted in the perihematomal area. Three random brain sections per slice were used for the mean under a microscopic field of 400x magnification by an independent observer. Data were calculated as the ratio of TUNEL-positive neurons (%) to total neurons.
3
Sperm DNA Fragmentation Analysis by TUNEL
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) technique, the In-Situ Cell Death Detection Kit with fluorescein (Roche Diagnostics GmbH, Mannheim, Germany) has been used according to Nur et al. [31 (link)]. Briefly, different aliquots of semen samples were diluted with phosphate-buffered saline (PBS) and centrifuged at 400 × g for 10 minutes. One drop of resuspended spermatozoa was smeared on a glass slide and fixed with 10% formaldehyde for 30 minutes at room temperature.
The slides were washed three times with PBS (5 minutes each), treated in a humidified chamber with proteinase K for 10 minutes, washed with PBS, treated with 3% H2O2 in distilled water for 10 minutes at room temperature, and washed again with PBS. The slide was permeabilized with 0.1% Triton X-100 for 5 minutes on ice. The slides were incubated in dark at 37°C for one hour with the TUNEL reaction mixture. After labeling, samples were washed with PBS and analyzed immediately using fluorescence microscopy (Zeiss Eurostar, Germany 100×). The percentage of TUNEL-positive sperm was determined by the evaluation of at least 100 sperm.
The slides were washed three times with PBS (5 minutes each), treated in a humidified chamber with proteinase K for 10 minutes, washed with PBS, treated with 3% H2O2 in distilled water for 10 minutes at room temperature, and washed again with PBS. The slide was permeabilized with 0.1% Triton X-100 for 5 minutes on ice. The slides were incubated in dark at 37°C for one hour with the TUNEL reaction mixture. After labeling, samples were washed with PBS and analyzed immediately using fluorescence microscopy (Zeiss Eurostar, Germany 100×). The percentage of TUNEL-positive sperm was determined by the evaluation of at least 100 sperm.
4
Tumor Cell Proliferation and Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tumor samples were evaluated by H&E staining, anti–Ki‐67, and TUNEL. Paraffin sections of tumor tissues were stained immunohistochemically using anti–Ki‐67 as the primary antibody and DAKO Envision Kit/HRP as the secondary antibody (DAKO). The TUNEL assay was performed using an In Situ Cell Death Detection Kit with fluorescein (Roche Diagnostics, Basel, Switzerland) according to the instructions provided by the manufacturer. The number of Ki‐67‐positive cells and TUNEL‐positive cells were counted in three random fields (×400 respectively) in three tumors.
5
Immunohistochemical Analysis of Rat Brain
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Staining of slices was performed as previously described (Wang et al., 2019 (link)). Briefly, rats were deeply anesthetized and perfused transcardially with ice-cold 4% paraformaldehyde. Rat's brain was post-fixed with 4% paraformaldehyde for 12 h, dehydrated with 30% sucrose/phosphate-buffered saline (PBS) at 4°C for 3 days, and then cut into 10-μm thickness coronal slices (−2.5 to −5 mm from bregma) using a Leica cryostat microtome. Coronal slices were permeabilized with 0.5% Triton X-100 and blocked with 5% goat serum, incubated with anti-NeuN rabbit antibody (1:200, #12943, CST), anti-NeuN mouse antibody (1:200, #94403, CST), anti-HSP27 (1:200, ab2790, Abcam), anti-Iba-1 (1:200, #19741, Wako) primary antibody at 4°C overnight and then incubated with the Anti-Mouse IgG-TRITC (1:100, T5393, Sigma) or Anti-Rabbit IgG-FITC (1:100, F9887, Sigma) at room temperature for 2 h. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining was assayed using in situ Cell Death Detection Kit with Fluorescein (11684795910, Roche) according to the manufacturer's instruction. Images were captured using a fluorescence microscope (BX51, Olympus) and analyzed with Image J software.
6
Quantifying Liver Apoptosis via TUNEL
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Liver apoptotic cells were quantitated by the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Cryopreserved liver tissue sections were cut at 5 µm on a cryomicrotome (Leica, Buffalo Grove, IL) and air-dried. TUNEL assay was performed using the In Situ Cell Death Detection Kit with Fluorescein (Roche Diagnostics, Indianapolis, IN) following the manufacturer's protocol. TUNEL-positive cells were counted using an inverted laser scanning confocal microscope (Zeiss LSM 780) with λem = 480 nm and λex = 525 nm. A total of five high-power fields (HPF) were analyzed for each animal.
7
Histopathological Analysis of Mouse Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All histopathological analysis was performed by an independent, certified pathologist at the Australian Phenomics Network, University of Melbourne. Monash and Sanger colonic and cecal sections were assessed using a scoring system based on previously established parameters (26 (link)), described in detail in Table S3 in the supplemental material. Terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) staining was performed on thymus sections using an in situ cell death detection kit with fluorescein (Roche), following the manufacturer’s instructions.
8
Quantifying Proliferation and Apoptosis in Lung Tissue
9
TUNEL Staining of Embryos
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole-mount TUNEL staining of developmentally staged control, morphant, and overexpression embryos was performed using the in situ cell death detection kit with fluorescein (Roche Applied Science, 11684795910). Embryos were then embedded in 1% low melting agarose and imaged with a Zeiss LSM780 confocal microscope and a 10× objective.
10
Assessing Cell Viability and Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell viability was assessed using the CellTiter 96 AQueous One Solution cell kit (Promega), which contains a tetrazolium compound MTS. Cl.8+ cells were seeded at a concentration of 1 × 106/ml in a flat-bottomed 96-well plate in minimal medium (MM) with or without recombinant IDGF2 (16 μg/ml), Ado and dAdo and incubated for 22 h. Next, 20 μl of MTS solution was added to each well and the plate was incubated for an additional 3 h at 25 °C. The absorbance at 490 nm was recorded with a 96-well plate reader. The assay was performed with five replicates for each sample.
Proliferation of Cl.8+ cells in SFM and CM was also measured by the direct counting of cells using digital photographs of identical areas (0.8 × 0.8 mm) taken every 24 hrs. Each value represents an average of three fields per plate.
TUNEL staining, BrdU incorporation, the ATP assay and the Ado uptake assay were performed as described previously20 (link). For TUNEL staining, we used the In Situ Cell Death Detection kit with fluorescein (Roche), and for BrdU incorporation, we used the In Situ Cell Proliferation Kit, FLUOS (Roche). ATP was measured by The CellTiter-Glo®Luminescent Cell Viability Assay (Promega). The Ado uptake assay was performed using H3-Ado in MM.
Proliferation of Cl.8+ cells in SFM and CM was also measured by the direct counting of cells using digital photographs of identical areas (0.8 × 0.8 mm) taken every 24 hrs. Each value represents an average of three fields per plate.
TUNEL staining, BrdU incorporation, the ATP assay and the Ado uptake assay were performed as described previously20 (link). For TUNEL staining, we used the In Situ Cell Death Detection kit with fluorescein (Roche), and for BrdU incorporation, we used the In Situ Cell Proliferation Kit, FLUOS (Roche). ATP was measured by The CellTiter-Glo®Luminescent Cell Viability Assay (Promega). The Ado uptake assay was performed using H3-Ado in MM.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!