Key features and details
- Assay type: Direct
- Detection method: Fluorescent
- Platform: Flow cytometer, Fluorescence microscope
- Assay time: 2 hr 30 min
- Sample type: Adherent cells, Suspension cells
Product nameTUNEL Assay Kit - FITC
See all DNA fragmentation kits
Sample typeAdherent cells, Suspension cells
Assay time2h 30m
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 37ºC
- 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 TUNEL Assay Kit - BrdU-Red (ab66110).
For chromogenic TUNEL staining, we recommend TUNEL Assay Kit - HRP-DAB ab206386.
Find out more about the TUNEL method in the TUNEL staining / TUNEL assay guide.
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
PlatformFlow cytometer, Fluorescence microscope
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 50 tests FITC-dUTP Orange 1 x 0.4ml Negative Control Cells Neutral 1 x 5ml PI/RNase Staining Buffer Amber bottle 1 x 25ml Positive Control Cells Brown 1 x 5ml Reaction Buffer Green 1 x 500µl Rinse Buffer Red 1 x 100ml TdT Enzymes 1 x 38µl Wash Buffer Blue 1 x 100ml
RelevanceInternucleosomal DNA fragmentation is a hallmark of apoptosis in mammalian cells.
- FCCP, mitochondrial oxidative phosphorylation uncoupler (ab120081)
- Cycloheximide, Protein synthesis inhibitor (ab120093)
- SB431542, ALK inhibitor (ab120163)
- Z-VAD(OMe)-FMK, Cell permeable, irreversible pan-caspase inhibitor (ab120487)
- Doxorubicin hydrochloride, Topoisomerase II inhibitor (ab120629)
- Dorsomorphin (Compound C), AMP-kinase inhibitor (ab120843)
- Calcein AM, fluorescent dye for cell viability (ab141420)
Voloboueva et al used TUNEL assay ab66108 to examine the effect of miR-210 inhibition on mitochondrial function and protection against apoptosis.
The reduced number of TUNEL+ve green cells in panel F indicates that the inhibition of miR-210 reduces the degree of apoptosis in cells treated with media preconditioned (CM) by proinflammatory activated microglia.
Green is TUNEL staining. Red is immunostaining of DCX protein. Representative images are shown.
Control RAW 264.7 cells.
RAW 264.7 cells treated with 2 μM camptothecin (ab120115) for 24 hours prior to staining.
RAW 264.7 cells treated with 10 μM camptothecin (ab120115) for 24 hours prior to staining.
ab66108 has been referenced in 22 publications.
- Tsao YC et al. Discovery of Isoplumbagin as a Novel NQO1 Substrate and Anti-Cancer Quinone. Int J Mol Sci 21:N/A (2020). PubMed: 32575541
- De Felice F et al. Sulodexide counteracts endothelial dysfunction induced by metabolic or non-metabolic stresses through activation of the autophagic program. Eur Rev Med Pharmacol Sci 23:2669-2680 (2019). PubMed: 30964194
- Li C et al. An investigation on the cytotoxicity and caspase-mediated apoptotic effect of biologically synthesized gold nanoparticles using Cardiospermum halicacabum on AGS gastric carcinoma cells. Int J Nanomedicine 14:951-962 (2019). PubMed: 30787609
- Umar SA et al. Glycyrrhizic Acid Prevents Oxidative Stress Mediated DNA Damage Response through Modulation of Autophagy in Ultraviolet-B-Irradiated Human Primary Dermal Fibroblasts. Cell Physiol Biochem 53:242-257 (2019). PubMed: 31313540
- Yu W et al. Yap overexpression attenuates septic cardiomyopathy by inhibiting DRP1-related mitochondrial fission and activating the ERK signaling pathway. J Recept Signal Transduct Res 39:175-186 (2019). PubMed: 31354091