Advantages of the Apo-ONE® Homogeneous Caspase-3/7 Assay:
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Convenient: The Homogeneous Caspase-3/7 Reagent is made simply by
diluting the substrate 1:100 with the assay buffer.
Fast: Minimal time is required for sample preparation and for generation
of statistically relevant data. One hour of incubation with the Homogeneous
Caspase-3/7 Reagent and cultured cells is typically sufficient, although longer
incubations are not detrimental and may benefit signal-to-noise ratios.
Flexible: Assay volume is flexible as long as the 1:1 ratio of reagent
volume to sample volume is preserved. The assay can be performed in a cuvette,
96 or 384 well plate. Furthermore, data can be collected within one hour and for
up to 18 hours after Reagent addition. |
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Sensitive: Detects significant caspase-3/7 activity in low numbers of
total cells (cell numbers in the hundreds). Longer incubations can be used to
obtain results in systems using low numbers of cells or in cells that express
low levels of caspase-3/7.
Informative: Defines mechanistic toxicity. The assay can be used to
conveniently assess apoptotic induction or inhibition after cell culture
treatment.
Automatable: The assay's "add-mix-read" format can be easily
integrated into automated platforms such as the Biomek® 2000 and
Biomek® FX
(Beckman). |
The Apo-ONE® Homogeneous Caspase-3/7 Assay is a revolutionary improvement in apoptosis detection methodology. Promega's proprietary
lysis/activity buffer, in conjunction with the (Z-DEVD)2-Rhodamine 110 substrate, enables a simple "add-mix-read" format for the detection of caspase-3 and -7 in adherent, suspension, and primary culture cells, or in purified caspase preparations. The homogeneous format eliminates tedious washing, concentration, and multiple freeze-thaw steps required by conventional caspase detection assays, resulting in a dramatic reduction in sample preparation time. The rhodamine 110-based substrate allows for exquisite sensitivity previously unobtainable with conventional colorimetric or fluorometric assays. Furthermore, the assay is inherently flexible for use in a variety of volumes and formats from cuvettes to 384 well plates.
Figures (click on a link to view a figure):
References
1. Learish, R.L., Bruss, M.D., and Haak-Frendscho, M. (2000) Dev. Brain Res.
122, 97–109.
2. Nicholson, D.W. et al. (1995) Nature 376, 37–43.
3. Garcia-Calvo, M. (1999) Cell Death Differ. 6, 362–9.
Footnotes:
Apo-ONE is a registered trademark of Promega Corporation. Biomek is a registered trademark of Beckman Instruments, Inc. Costar is a registered trademark of Corning, Inc. Cytofluor is a registered trademark of Perseptive
Biosystems, Inc. Microtiter is a registered trademark of DYNEX Technologies, Inc. Prism is a registered trademark of E.I. du Pont de Nemours and Company, Inc.
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