Applications

• Detect fragmented nucleosomal DNA
• Screening for apoptotic cells
• Fluorescence microscopy
• Flow cytometry

Benefits

• High sensitivity
• Appropriate for various multiwell formats and automation
• Compatible with standard lab equipment

The PromoKine DNA Fragmentation Detection Kits I and II are based on the TUNEL method and identify DNA strand breaks occuring during apoptosis. The kits provide all required components - including positive and negative control cells and tissue slides - for convenient detection of DNA fragmentation by fluorescence microscopy or flow cytometry. The TUNEL assay employs a terminal deoxynucleotidyl transferase (TdT) to add Br-dUTP (bromolated deoxyuridine triphosphate nucleotides) to the fragmented DNA strands. Br-dUTP is more readily incorporated into DNA strand breaks than other larger ligands (e.g., fluorescein, biotin or digoxigenin). The greater incorporation gives rise to a brighter signal when the Br-dUTP sites are identified by a green-fluorescent (Kit I) or red-fluorescent (Kit II) labeled anti-BrdU monoclonal antibody. The labeled DNA can then be observed by fluorescence microscopy or analyzed by flow cytometry. The DNA Fragmentation Kit II works also very well with GFP transfected cells.

The Fluorometric DNA Fragmentation Detection Kit III is a modified TUNEL assay and provides all reagents required for the specific detection of apototic cells within a cell population in tissue sections and fixed cell preps. The kit utilizes terminal deoxynucleotidyl transferase (TdT) to catalyze incorporation of fluorescein-12-dUTP at the free 3'-hydroxyl ends of the fragmented DNA. The fluorescein-labeled DNA can then be observed by fluorescence microscopy or analyzed by flow cytometry.

The ready-to-use, TUNEL-based Colorimetric DNA Fragmentation Detection Kit (IHC) has been specifically designed for convenient detection of DNA fragmentation in tissue sections and fixed cell preps. It includes positive and negative control slides and utilizes Br-dUTP, biotin-labeled anti-BrdU antibodies and HRP-streptavidin conjugate. Immunohistochemical staining results can be microscopically analyzed.

The Apoptotic DNA Ladder Detection Kit I provides an easy and sensitive means for detecting DNA fragmentation in apoptotic cells. The method requires less than 90 minutes to prepare the DNA, without the need for extraction or columns. DNA fragmentation can then easily be visualized by agarose gel electrophoresis. The procedure increases recovery of small fragmented DNA, and therefore improves the sensitivity of the assay. The assay can be used to detect apoptotic DNA ladders, extracting both genomic DNA and laddered DNA.

Using the Apoptotic DNA Ladder Extraction Kit II allows to selectively isolate DNA ladders without interference of regular genomic DNA. Apoptotic DNA ladders from both tissues and cells with as little as 5% or less apoptotic cell population can be detected. The kit specifically extracts laddered DNA, not genomic DNA.