Product nameTUNEL Assay Kit - FITC
See all DNA fragmentation kits
Sample typeAdherent cells, Suspension cells
Assay time2h 30m
TUNEL Assay Kit - FITC ab66108 (previously called TUNEL Assay Kit - In situ Direct DNA Fragmentation) provides complete components including positive and negative control cells for conveniently detecting DNA fragmentation by fluorescence microscopy or flow cytometry.
The TUNEL assay protocol uses terminal deoxynucleotidyl transferase (TdT) to catalyze the incorporation of fluorescein-12-dUTP at the free 3'-hydroxyl ends of the fragmented DNA. The fluorescein-labeled DNA can then be analyzed by flow cytometry or fluoresence microscopy with Ex/Em 488/520 nm. Propidium iodide is also included in the 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).
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.
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.
This product has been referenced in:
- Azam F et al. Fagonia indica Repairs Hepatic Damage through Expression Regulation of Toll-Like Receptors in a Liver Injury Model. J Immunol Res 2018:7967135 (2018). Read more (PubMed: 30057922) »
- Sun X et al. Valsartan prevents glycerol-induced acute kidney injury in male albino rats by downregulating TLR4 and NF-?B expression. Int J Biol Macromol 119:565-571 (2018). Read more (PubMed: 30053391) »