Description
Key facts
Detection method:Fluorescent,
Sample types:Suspension cells, Adherent cells,
Assay type:Direct,
Assay time:2h 30m,
Assay Platform:Flow cytometer, Fluorescence microscope
Product details:
How the assay works
TUNEL assay - FITC ab66108 is used to detect DNA fragmentation in apoptosis.
The TUNEL assay - FITC used to detect DNA fragmentation, such as in apoptosis. It 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.
The TUNEL assay protocol in this kit uses the deoxynucleotide fluorescein-12-dUTP. DNA which has been labeled with fluorescein can then be analyzed by flow cytometry or fluoresence microscopy with Ex/Em 488/520 nm. Propidium iodide is also included in this kit as a counterstain with Ex/Em 488/623 nm.
TUNEL assay protocol summary:
- fix cells with formaldehyde for 15 min on ice
- wash with PBS
- add ice-cold 70% ethanol and incubate for 30 min
- pellet cells and resuspend in wash buffer and wash again
- pellet cells and resuspend in staining solution and incubate for 60 min at 37°C
- add rinse buffer, pellet cells and discard supernatant, and rinse again
- resuspend cells in propidium iodide/RNAse A solution and incubate for 30 min at room temp
- analyze with flow cytometer or fluoresence microscope
To use HRP detection, we recommend TUNEL Assay Kit - HRP-DAB
ab206386
. To use BrdU-Red (Ex/Em = 488/576nm) as a label, we recommend
ab66110
How other researchers are using
TUNEL Assay Kit - FITC has been used in variety of sample type including:
- Human lung cancer cell lines A549 and Human colon cancer cell lines HCT-116 1
- Mouse lung tissue
- Mouse Mesenchymal stem cell (MSC) 3
References:
1 - Olszewski M et al. 2024; 2 - Zhang P et at 2024; 3 - Zhang T et al. 2023
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
