Description
Key facts
Detection method:Colorimetric,
Sample types:Tissue, Adherent cells,
Assay type:Cell-based (qualitative),
Assay time:5h
Product details:
TUNEL Assay Kit ab206386 is used to detect DNA fragmentation, such as in apoptosis. TUNEL assay kits are sometimes also called In Situ Cell Death Detection Kits.
How the TUNEL assay works
The TUNEL assay exploits the fact that apoptotic endonucleases not only affect cellular DNA by producing the classical DNA ladder, but also generate free 3’-OH groups at the ends of these DNA fragments. These free 3’-OH groups are end-labeled in the TUNEL assay allowing for the detection of apoptotic cells using a molecular biology-based, end labeling technique.
The TUNEL assays uses terminal deoxynucleotidyl transferase (TdT) to catalyze the incorporation of deoxynucleotides at the free 3'-hydroxyl ends of fragmented DNA. The deoxynucleotides are then labeled in a variety of ways for detection of the degree of DNA fragmentation.
TUNEL assay protocol summary
In this TUNEL assay protocol:
- Terminal deoxynucleotidyl Transferase (TdT) binds to exposed 3'-OH ends of DNA fragments generated in response to apoptotic signals and catalyzes the addition of biotin-labeled deoxynucleotides.
- Biotinylated nucleotides are bound with a streptavidin-horseradish peroxidase (HRP) conjugate.
- Diaminobenzidine (DAB) reacts with the HRP labeled sample to generate an insoluble colored (brown) substrate at the site of DNA fragmentation.
- Counterstaining with methyl green aids in the evaluation of normal and apoptotic cells.
Related TUNEL assay products
This kit is designed for chromogenic TUNEL staining with HRP and DAB. To use FITC (Ex/Em = 495/519 nm) as a label, we recommend
ab66108
. To use BrdU-Red (Ex/Em = 488/576nm) as a label, we recommend
ab66110
How other researchers are using TUNEL assay kit ab206386
This TUNEL assay kit has been cited in several publications in a variety of sample types, including:
- Human: Precision-cut intestinal slices (hPCIS)
(paraffin-embedded sections), human gastric cancer cell culture
(single cell suspension)
- Mouse: abdominal aortic aneurysm lesion
, thyroid gland
, lung
, heart
, pancreas
, ovary
, skin
, liver
(paraffin-embedded sections), brain
(frozen sections)
- Rat: heart
, brain
(paraffin-embedded sections)
- Other: human xenograft tumours in mice
, tissue from nude mice injected with human pancreatic cancer cell line
, pig heart
(paraffin-embedded sections)
References: 1- PMID: 35428896; 2- PMID: 34440988; 3- PMID: 30651000; 4- PMID: 30952933; 5- PMID: 40084179; 6- PMID: 31886717; 7- PMID: 36458554; 8- PMID: 29379893, PMID: 39469582; 9- PMID: 33690222; 10- PMID: 36911196; PMID: 35970323; PMID: 35763355; 11- PMID: 34518549; 12- PMID: 34543287; PMID: 32536996; 13- PMID: 38413663; 14- PMID: 35265825; 15- PMID: 30755605; 16- PMID: 32784904
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
