Science Catalog 2012 Life Cell Signaling DeadEnd™ Fluorometric TUNEL System Product DeadEnd™ Fluorometric TUNEL System For Research Use Only. Not for Use in Diagnostic Procedures. Description: The DeadEnd™ Fluorometric TUNEL System is a classic TUNEL Assay designed for the specific detection and quantitation of apoptotic cells within a cell population. The DeadEnd™ Fluorometric TUNEL System measures nuclear DNA fragmentation, an important biochemical hallmark of apoptosis in many cell types. The system is nonradioactive and provides simple, accurate and rapid detection of apoptotic cells in situ at the single-cell level or in cell suspensions. The DeadEnd™ Fluorometric TUNEL System measures the fragmented DNA of apoptotic cells by catalytically incorporating fluorescein12-dUTP at 3´-OH DNA ends using the enzyme Terminal Deoxynucleotidyl Transferase (TdT), which forms a polymeric tail using the principle of the TUNEL (TdT-mediated dUTP Nick-End Labeling) assay. The fluorescein-12-dUTPlabeled DNA can then be visualized directly by fluorescence microscopy or quantitated by flow cytometry. Features: • Save Money: System provides sufficient reagents for 60 assays of 50μl each. • Save Time: Direct incorporation of fluorescent nucleotide reduces number of incubation steps. • Choose Sample Type: Use to detect apoptosis in cultured cells and formalin-fixed, paraffin-embedded tissue sections. • Convenient: Plastic coverslips provided simplify sample handling. Storage Conditions: Store at –20°C. Store the Nucleotide Mix protected from light at –20°C. Protocol DeadEnd™ Fluorometric TUNEL System Technical Bulletin For Research Use Only. Not for Use in Diagnostic Procedures. Description: Z-VAD-FMK (carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]fluoromethylketone) is a cell-permeant pan caspase inhibitor that irreversibly binds to the catalytic site of caspase proteases and can inhibit induction of apoptosis. For inhibition of apoptosis, Z-VAD-FMK should be added at the same time that apoptosis is induced. Z-VAD-FMK is provided at 20mM in DMSO for convenient addition to cell culture or extracts. The peptide is O-methylated in the P1 position on aspartic acid, providing enhanced stability and increased cell permeability. The suggested concentration for use in the anti-Fas mAb-treated Jurkat cell culture model system is 20μM. Storage Conditions: Store at –20°C protected from light and moisture. Caspase Inhibitor Ac-DEVD-CHO Product Caspase Inhibitor Ac-DEVD-CHO Size Conc. Cat.# Price (Fr) 100 µl 10 mM G5961 349.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: Ac-DEVD-CHO is an inhibitor of caspase-3/7 (DEVDase) activity. The concentration of inhibitor required to inhibit caspase activity must be determined empirically for each system. Ten micromolar inhibitor is sufficient to inhibit caspase activity in extracts of apoptotic THP-1 cells. Ac-DEVD-CHO is supplied as a 10mM solution in DMSO. Part# TB235 Storage Conditions: Store at –20°C protected from light and moisture. Digitonin Product Digitonin For Research Use Only. Not for Use in Diagnostic Procedures. Description: Digitonin is a detergent solution useful for permeabilizing cells and for creating a cytotoxicity chemistry positive control. Storage Conditions: Store at -20°C protected from light. Size Cat.# Price (Fr) 40 µl G9441 Pls. Enq. Size Cat.# Price (Fr) 60 reactions G3250 891.00 Caspase Inhibitor Z-VAD-FMK Product Caspase Inhibitor Z-VAD-FMK, 20mM Size Cat.# Price (Fr) 50 µl G7231 497.00 125 µl G7232 870.00 Neural progenitor cells migrating away from a spherical cluster of apoptotic cells. The condensed nuclei (green) contain fragmented DNA, as indicated by fluorescent labeling with the DeadEnd™ Fluorometric TUNEL System, in contrast with larger intact nuclei stained with DAPI (blue). The cells were also processed for immunocytochemical staining using a primary antibody to βIII Tubulin (Cat.# G7121) and a Cy®3-conjugated secondary antibody where immature processbearing neurons (red) are distinctly labeled. 32 For complete and up-to-date product information visit: www.promega.com/catalog 2272CA06_8A Pagina 35
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