Cell Signaling CaspACE™ Assay System, Colorimetric Product CaspACE™ Assay System, Colorimetric For Laboratory Use. Description: The CaspACE™ Assay System, Colorimetric, provides reagents for measuring caspase-3 activity. The system includes a colorimetric substrate and a cell-permeant inhibitor that allow quantitative measurement of caspase-3 (DEVDase) protease activity. The colorimetric substrate (Ac-DEVD-pNA) is labeled with the chromophore p-nitroaniline (pNA). pNA is released from the substrate upon cleavage by DEVDase. Free pNA produces a yellow color that is monitored by a spectrophotometer at 405nm. The amount of yellow color produced is proportional to the amount of DEVDase activity present in the sample. The potent, irreversible and cell-permeant pan-caspase inhibitor Z-VAD-FMK is provided in the CaspACE™ Assay System, Colorimetric. The addition of the Z-VAD-FMK Inhibitor prior to the induction of apoptosis in cell culture inhibits the activation of the caspase cascade, including caspase-3. Features: • Timely: Measures an early indicator of apoptosis. • Quantitative or Qualitative: Determine total caspase-3 activity or screen for inducers or inhibitors of caspase activity. • Versatile: May be used with purified enzyme preparations, cell extracts or tissue lysates. Storage Conditions: Store at –20°C. Store substrates and inhibitors in aliquots at –20°C away from light and moisture. Protocol CaspACE™ Assay System, Colorimetric Technical Bulletin CaspACE™ FITC-VAD-FMK In Situ Marker Product CaspACE™ FITC-VAD-FMK In Situ Marker For Research Use Only. Not for Use in Diagnostic Procedures. Description: CaspACE™ FITC-VAD-FMK In Situ Marker is a fluorescent analog of the pan caspase inhibitor Z-VAD-FMK (carbobenzoxy-valyl-alanylaspartyl-[O-methyl]-fluoromethylketone). The fluorescein isothiocyanate (FITC) group has been substituted for the carbobenzoxy (Z) N-terminal blocking group to create the fluorescent apoptosis marker. This structure allows delivery of the inhibitor into the cell where it irreversibly binds to activated caspases. The FITC label enables a single-reagent addition to assay for caspase activity in situ. The FITC-VAD-FMK is supplied as a 5mM solution in DMSO and is intended for in situ monitoring of caspase activity by fluorescence detection. The suggested concentration for use in anti-Fas-treated Jurkat cell culture is 10μM. Features: • Simplify Your Protocol: Add FITC-VAD-FMK, incubate, wash and view fluorescence. No preparation of cell extracts or long incubations. • Use a Variety of Detection Methods: Detect apoptotic cells by microscopy or flow cytometry; combine with other markers to assess cell populations or determine apoptotic frequency within a population; adaptable to high-throughput applications. • Get Reliable Results: Synthesized peptide provides consistent results from every batch, unlike Annexin V, which can be highly variable between batches. • Use With Live Cells: Easily moves in and out of cells and remains anchored inside cultured apoptotic cells. Storage Conditions: Store at –20°C protected from light and moisture. Protocol CaspACE™ FITC-VAD-FMK In Situ Marker Protocol Part# 9PIG746 For complete and up-to-date product information visit: www.promega.com/catalog 31 Size Cat.# Price (Fr) 50 µl G7461 473.00 125 µl G7462 1006.00 Part# TB270 Size Cat.# Price (Fr) 50 assays G7351 594.00 100 assays G7220 891.00 DeadEnd™ Colorimetric TUNEL System Product DeadEnd™ Colorimetric TUNEL System For Research Use Only. Not for Use in Diagnostic Procedures. Description: The DeadEnd™ Colorimetric TUNEL System is a modified TUNEL Assay designed to provide simple, accurate and rapid detection of apoptotic cells in situ at the single-cell level. The DeadEnd™ Colorimetric TUNEL System measures nuclear DNA fragmentation, an important biochemical indicator of apoptosis. The system can be used to assay apoptotic cell death in tissue sections and cultured cells. The DeadEnd™ Colorimetric TUNEL System end-labels the fragmented DNA of apoptotic cells using a modified TUNEL (TdT-mediated dUTP Nick-End Labeling) assay. Biotinylated nucleotide is incorporated at the 3´-OH DNA ends using the enzyme Terminal Deoxynucleotidyl Transferase (TdT). Horseradish-peroxidase-labeled streptavidin (Streptavidin HRP) is then bound to these biotinylated nucleotides, which are detected using the peroxidase substrate, hydrogen peroxide, and the stable chromogen, diaminobenzidine (DAB). Using this procedure, apoptotic nuclei are stained dark brown. Features: • Assay Cells or Tissue: Detect apoptosis in thick tissue sections or assess cell morphology. • Simplify: Includes DAB substrate and H2O2 for color detection and plastic coverslips that simplify sample handling. • Proven Applications: Vibratome® sections of neuronal tissue, Jurkat cells, HL-60 cells. Storage Conditions: Store the Equilibration Buffer, TdT Enzyme, Biotinylated Nucleotide Mix and Proteinase K at –20°C. Store the Streptavidin HRP, DAB 20X Chromogen, DAB Substrate 20X Buffer and Hydrogen Peroxide 20X at 4°C. Store the SSC 20X and Plastic Coverslips at room temperature. Protocol DeadEnd™ Colorimetric TUNEL System Technical Bulletin Part# TB199 Size Cat.# Price (Fr) 20 reactions G7360 535.00 40 reactions G7130 803.00 2 Cell Health Assays Pagina 34
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