Description
Key facts
Detection method:Fluorescent,
Sample types:Tissue, Suspension cells, Adherent cells,
Assay type:Quantitative,
Assay time:3h,
Assay Platform:Flow cytometer, Fluorescence microscope
Product details:
TUNEL Assay Kit - BrdU-Red (ab66110) is a sensitive DNA fragmentation detection kit using Br-dUTP for enhanced signal detection.
How the assay works
The TUNEL assay 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.
This TUNEL assay protocol is based on Br-dUTP (bromolated deoxyuridine triphosphate nucleotide), which can be more readily incorporated into DNA strand breaks by the TdT enzyme than other dUTP labels such as FITC, biotin or dioxigenin. The greater incorporation rate produces a brighter signal when the Br-dUTP sites are detected with an anti-BrdU monoclonal antibody directly labeled with a red fluorochrome.
The BrdU-Red signal can be analyzed at Ex/Em 488/576 nm, with an optional 7-AAD counterstain at Ex/Em 488/655nm.
This TUNEL assay kit includes both negative and positive control cells. It is designed to be suitable for studying DNA fragmentation in GFP-transfected cells.
Tunel assay kit BrdU-Red protocol summary:
- fix cells / tissues with formaldehyde, or deparaffinize and rehydrate if paraffin sections, and wash
- incubate cells in 70% ethanol for 30 min at 4°C, or if tissues incubate with proteinase K solution for 5 min at room temp and refix with formaldehyde
- wash
- incubate in DNA labeling solution for 60 min at 37°C
- wash
- incubate in antibody solution for 30 min at room temp
- add 7-AAD / RNase A solution and incubate for 30 min at room temp
- analyze with flow cytometry or fluorescent microscopy
To use HRP detection, we recommend TUNEL Assay Kit - HRP-DAB
ab206386
. To use FITC (Ex/Em = 495/519 nm) as a label, we recommend TUNEL Assay Kit - FITC
ab66108
How other researchers are using
TUNEL Assay Kit - BrdU-Red has been used in variety of sample type including:
- Mouse myocardial tissues 1
- Human salivary gland tissues 2
- Mouse fibroblasts
References:
1 -Qi P et al. 2023; 2 - Nakamura H et at 2023; 3 - Kawamura Y et al. 2023.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
