TUNEL Assay Kit - FITC

TUNEL Assay Kit - FITC ab66108 provides complete components, including positive and negative control cells, for detecting DNA fragmentation by fluorescence microscopy or flow cytometry.

The TUNEL assay is 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

This kit is FITC-labeled (Ex/Em = 495/519nm). It was previously called TUNEL Assay Kit - In situ Direct DNA Fragmentation. To use BrdU-Red (Ex/Em = 488/576nm) as a label, we recommend . For chromogenic TUNEL staining, we recommend . Find out more about the TUNEL method in the . **How other researchers have used FITC TUNEL Assay Kit ab66108** This TUNEL assay kit has been used in publications in a variety of sample types, including: - Human: HUVEC cell cultures1, AGS gastric carcinoma cells by imaging2, gastric tumor cell xenograft cells by flow cytometry3, MDA-MB-231 breast cancer xenograft tissue sections by imaging4, neural blastoma cell cultures by flow cytometry5, SH-SY5Y cells by flow cytometry6, A549 cells by flow cytometry7, Huh7 and HepG2 cell cultures by imaging8- Mouse: liver tissue by imaging9, cultured neural stem cells by imaging10 - Rat: kidney tissue sections by imaging11 References: 1 - De Felice F et al 2019, 2 - Li C et al 2019, 3 - Lau WM et al 2018, 4 - Chung SJ et al 2017, 5 - Sobham PK et al 2017, 6 - Albarran L et al 2016, 7 - Lamb SA et al 2014, 8 - Fu B et al 2016, 9 - Azam F et al 2018, 10 - Voloboueva LA et al 2017, 10 - Sun X et al 2015, 11 - Chen J et al 2015