Homogeneous caspase-3/7 assay for the detection of apoptosis combines extraordinary sensitivity with unparalleled speed and simplicity
Promega's Apo-One homogeneous caspase-3/7 assay for the detection of apoptosis combines extraordinary sensitivity with unparalleled speed and simplicity.
Developed with automation in mind, results can be obtained in as little as an hour.
Apo-One is the first apoptosis system with an ultra-simple 'add, mix, read' format.
No separate extraction step is required.
With Apo-One laboratories can precisely and rapidly monitor toxic effects or disease symptoms in both small-scale or HTS systems. "The range of diseases in which apoptosis has been implicated (cancer, heart attacks, stroke, neurodegenerative disease) emphasises the importance of this new assay," comments Derek McCall, managing director, Promega UK.
"The Apo-One assay represents a revolutionary improvement in apoptosis detection methodology." Promega's proprietary lysis/activity buffer, in conjunction with the (Z-DevD)2-Rhodamine 110 substrate, eliminates the tedious washing, concentration and multiple freeze-thaw steps required by conventional caspase detection.
Bi-functional buffering ensures rapid and efficient lysis, while simultaneously supporting optimal caspase-3/7 enzyme activity.
Apoptosis can now be detected in adherent, suspension or primary cells in culture, with a sensitivity of a few hundred cells per well.
Activity or inhibition of purified caspase-3/7 enzymes can also be determined.
As fewer cells are required for accurate detection, savings can be made on cell culture reagents.
Assays can be performed in cuvettes, 96- or 384-well plates.
Assay volume is also scalable, as long as a 1:1 ratio of homogeneous caspase-3/7 reagent volume to culture volume is maintained.
For increased sensitivity, longer incubations can be used to obtain results from low numbers of cells or low levels of caspase-3/7 activity. A further advantage of the Apo-One assay is that it is possible to multiplex the rhodamine110 fluor molecule with other non-interfering fluorescent biological indicators for the detection of additional redox reactions.
