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In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (ab66110) is a convenient and sensitive method to detect DNA fragmentation by flow cytometry and fluorescence microscopy in live cells. The kit uses Br-dUTP (bromolated deoxyuridine triphosphate nucleotide), which can be more readily incorporated into DNA strand breaks that other dUTP labels such as fluorescein, 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 kit includes cells that can be used as positive or negative control. This assay is especially suitable for studying DNA fragmentation in GFP-transfected cells.
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This kit is BrdU-Red labeled (Ex/Em = 488/576 nm). If want to use FITC (Ex/Em = 495/519 nm) as a label, we recommend our In situ Direct DNA Fragmentation (TUNEL) Assay Kit (ab66108).
|7-AAD/RNase Staining Buffer||Amber bottle||1 x 30ml|
|Anti-BrdU-Red Antibody||Orange||1 x 300µl|
|Br-dUTP||Violet||1 x 480µl|
|Negative Control Cells||Neutral||1 x 5ml|
|Positive Control Cells||Brown||1 x 5ml|
|Reaction Buffer||Green||1 x 600µl|
|Rinse Buffer||Red||1 x 120ml|
|TdT Enzymes||1 x 45µl|
|Wash Buffer||Blue||1 x 120ml|
Our Abpromise guarantee covers the use of ab66110 in the following tested applications.
|FM||Use at an assay dependent concentration.|
|Flow Cyt||Use at an assay dependent concentration.|
Detection of DNA fragmention (TUNEL staining) using the negative and positive control cells (HL-60 untreated and treated with camptothecin). Cells were stained following the assay protocol. The fluorescence signal was detected and analyzed using BD FACScan System (Becton Dickinson).
TUNEL staining in whole mount Hydractinia echinata using In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (ab66110).
Animals were fixed in 4% PFA in PBS for 1 hour and processed as per the protocol without proteinaseK treatment. In place of proteinaseK animals were permeabilised in 3% Triton in PBS for 15 minutes. Animals were counter-stained with DAPI.