The slices were routinely dewaxed, hydrated, and transferred into a wet box. Proteinase K (50 µg/ml) solution was added and incubated at 37 °C for 30 min. Then, they were washed with PBS for three times (5 min each time). TUNEL test solution (Beyotime Biotechnology) was added and incubated at 37 °C in dark for 1 h. The slices were mounted and observed under the fluorescence microscope (CKX53, Olympus Corp.).
Tunel test solution
Manufactured by Beyotime
Sourced in China
The TUNEL test solution is a laboratory reagent used for the detection of DNA fragmentation, a hallmark of apoptosis or programmed cell death. The solution contains the necessary components to perform the TUNEL (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) assay, a widely used technique for identifying and quantifying cells undergoing apoptosis.
Automatically generated - may contain errors
6 protocols using tunel test solution
1
TUNEL Assay for Apoptosis
2
Assessing Caspase Activity and Apoptosis in TM-4 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A Caspase 3/8/9 Activity Assay Kit (Beyotime, China) was used to assess caspase 3/8/9 activity. TM-4 cells in 6-well plates were treated with TiO2-NPs for 24 h and then digested with trypsin for 2 min. Total proteins were extracted using RIPA buffer (Beyotime, China). The supernatants were collected by centrifugation at 13,000 rpm for 15 min at 4° C. The test buffer, the samples, and Ac-DEVD/IETD/LEHD-pNA (2 mM) were added to 96-well plates and incubated at 37° C for 2–4 h or overnight. Then, OD405 was measured. Protein concentrations were measured using the Bradford method to calculate the units of caspase 3/8/9 activity per sample based on the units of weight protein.
For the TUNEL apoptosis assay, TM-4 cells were fixed in 4% PFA for 30 min and permeabilized in 0.3% Triton X-100 for 5 min. Then, samples were incubated with TUNEL test solution (Beyotime, China) for 1 h at 37° C. Nuclei were stained blue with DAPI (Beyotime, China). After mounting with Antifade Mounting Medium (Beyotime, China), the samples were detected using a confocal laser-scanning microscope (CLSM 510) (Carl Zeiss, Germany).
For the TUNEL apoptosis assay, TM-4 cells were fixed in 4% PFA for 30 min and permeabilized in 0.3% Triton X-100 for 5 min. Then, samples were incubated with TUNEL test solution (Beyotime, China) for 1 h at 37° C. Nuclei were stained blue with DAPI (Beyotime, China). After mounting with Antifade Mounting Medium (Beyotime, China), the samples were detected using a confocal laser-scanning microscope (CLSM 510) (Carl Zeiss, Germany).
3
TUNEL Assay for Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transfected cells were washed with PBS and fixed with 4% paraformaldehyde for 30 minutes. Subsequently, cells were incubated with PBS containing 0.3% Triton X-100 for 5 minutes at room temperature. After washing with PBS, 50 µL TUNEL test solution (Beyotime, Shanghai, China) was added and incubated in the dark at 37 ℃ for 60 minutes. Cells were observed under the fluorescence microscope (IX53, Olympus, Japan). Each group was repeated three times.
4
Artemisinin-induced Apoptosis in SH-SY5Y Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SH-SY5Y cells were pre-treated with 5 μM Compound C (AMPK inhibitor) for 30 min, and treated with 12.5 μM artemisinin for 2 h, then incubated with or without 600 μM H2O2 for a further 24 h. After treatment, the cells were fixed with 4% PFA for 30 min and washed with 0.1% Triton-X PBS for 3 times. Then cells were incubated with TUNEL test solution (C1086, Beyotime, China) at 37 °C for 60 min in the dark following the manufacturer’s instruction. After washing with 1× PBS, images were taken with a fluorescence microscope.
5
TUNEL Assay for Apoptosis Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen sections were washed twice with PBS for 3 min each time and incubated in PBS containing 0.5% Triton™ X-100 for 5 min at room temperature. The incubated tissue sections were washed twice with PBS for 3 min each time. The sections were further incubated with 50 μl TUNEL test solution (Beyotime; China) at 37 °C for 1 h in dark. After incubation, they were washed with PBS three times for 3 min each time and observed under a fluorescence microscope. The excitation and emission wavelengths were 550 and 570 nm, respectively.
6
Quantifying Hepatocyte Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis was detected through the terminal deoxynucleotidyl transferase-dUTP nick-end labeling (TUNEL) assay or via PI/annexin V staining. For the TUNEL assay, frozen liver sections were permeabilized and incubated with the TUNEL test solution (Beyotime) for 60 min. Fluorescence was captured using a laser confocal microscope. For PI/annexin V staining, cells were resuspended in 100 μl of binding buffer containing 5 μl of PI and 10 μl of annexin V solution (BD Biosciences, USA) and incubated at 25℃ for 15 min. The number of apoptotic cells was measured via flow cytometry (ACEA NovoCyte, USA).
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!