Highly sensitive, homogeneous fluorescent apoptosis assay for use in cultured cells and with purified caspase. Just add-mix-read.

• Fast and Sensitive Measurement of Active Caspase-3 and -7
• Fluorescent "add-mix-read" apoptosis assay for use in cultured cells and with purified caspase
• Dramatically decrease time to results with highly sensitive and simple protocol
• Flexible assay design allows for adaptable formats and throughput needs

The Apo-ONE Homogeneous Caspase-3/7 Assay provides the necessary reagents for fast and sensitive measurement of active caspase-3 and -7 in a homogeneous format. The assay includes a profluorescent caspase-3/7 consensus substrate, rhodamine 110 bis-(N-CBZ-L-aspartyl-L-glutamyl-L-valyl-aspartic acid amide) (Z-DEVD-R110), and an optimized bifunctional cell lysis/activity buffer. The buffer efficiently lyses cultured mammalian cells and supports optimal caspase-3/7 enzymatic activity. The substrate and buffer are combined to make the Apo-ONE Caspase-3/7 Reagent that is added directly to samples. Upon cleavage on the C-terminal side of the aspartate residue in the DEVD peptide substrate sequence by caspase-3/7 enzymes, the rhodamine 110 becomes fluorescent when excited at a wavelength of 498nm. The emission maximum is 521nm. The amount of fluorescent product generated is representative of the amount of active caspase-3/7 present in the sample.

DeadEnd™ Colorimetric TUNEL System provides accurate detection of apoptotic cells in situ. It includes 600 units of recombinant terminal Deoxynucleotidyl transferase, 20 µL of biotinylated nucleotide mix, 4.8 mL of equilibration buffer, 40 µL of Streptavidin HRP, 200 µL of DAB substrate 20X buffer, 200 µL of hydrogen peroxide 20X, 200 µL of DAB 20X chromogen, 20 plastic coverslips, 20 mL of SSC 20X, and 10 mg Proteinase K. DeadEnd Fluorometric TUNEL System provides rapid detection of apoptotic cells in situ and can be used in cultured cells or tissue sections and includes three vials of 600 units of recombinant terminal Deoxynucleotidyl transferase, six vials of 50 µL nucleotide mix, 10 mg of proteinase K, 70 mL of SSC 20X, 60 plastic coverslips, and 9.6 mL equilibration buffer.

The assay includes a profluorescent caspase-3/7 consensus substrate and buffer for cell lysis and caspase-3/7 enzymatic activity. It works with purified enzyme preparations or cultured mammalian cells. CaspACE™ FITC-VAD-FMK In Situ Marker detects apoptotic cells with fluorescent microscopy or flow cytometry. The FITC-VAD-FMK marker structure allows delivery into cells to bind to activated caspases. Caspase-Glo™ Assay Systems are luminescent assays that measure caspase-3/7 activity or caspase 8 or 9 activity. The protocols can be applied to purified enzymes or cultured cells in multiwell plates.