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| Product Name | Caspase-Glo 3/7 Luminescence Assay Reagent |
| Catalog No. | CCAT-HMM-0046 |
| Description | A homogeneous bioluminescent reagent that measures caspase-3 and caspase-7 activities in cultured cells or purified enzyme preparations using a proluminescent substrate containing the DEVD tetrapeptide sequence. Upon caspase cleavage, the released aminoluciferin serves as a substrate for thermostable luciferase, generating a glow-type luminescent signal proportional to caspase activity. |
| Intended Use | High-throughput screening of caspase-3/7 activation in apoptosis assays, compound library screening for apoptosis modulators, and dose-response profiling of pro-apoptotic therapeutic candidates. |
| Principle / Technology | The luminogenic substrate Z-DEVD-aminoluciferin is cleaved specifically by caspase-3 and caspase-7 at the DEVD recognition site; the released aminoluciferin then reacts with Ultra-Glo recombinant luciferase, ATP, magnesium, and oxygen to produce oxyluciferin and light with a stable glow half-life exceeding 2 hours. |
| Detection Method | Bioluminescence detection with luminometer or multi-mode plate reader in luminescence mode; relative light units (RLU) proportional to caspase activity |
| Sample Type | Cultured mammalian cells in 96-well or 384-well opaque white-walled plates, purified recombinant caspase-3/7, cell and tissue lysates |
| Performance Range / Specifications | Linear caspase response: 0.01–10 ng purified caspase-3 per well; signal half-life >2 hours enabling batch plate processing; RLU range 100–10,000,000 |
| Sensitivity / LOD | Detection of caspase-3/7 activity from approximately 10–50 apoptotic cells per well under optimal conditions |
| Specificity | DEVD recognition sequence provides high specificity for caspase-3 and caspase-7; negligible cross-reactivity with caspase-8 or caspase-9; luciferase product non-lytic at standard concentration, maintaining cell membrane integrity for extended incubation |
| Reaction Conditions / Protocol | Equilibrate Caspase-Glo 3/7 reagent to room temperature; add equal volume of reagent to cell culture medium (e.g., 100 μL reagent to 100 μL medium in 96-well plate); mix contents gently on plate shaker at 300–500 rpm for 30 seconds; incubate 30 minutes to 3 hours at room temperature; measure luminescence with 0.5–1 second integration time per well |
| Components / Formulation | Single-component reagent: Z-DEVD-aminoluciferin proluminescent substrate, Ultra-Glo recombinant thermostable luciferase, ATP, MgSO4, proprietary cell lysis detergent, and buffer stabilizers in a proprietary lyophilized cake format; reconstitution buffer provided |
| Storage Conditions | Lyophilized reagent: -20°C protected from light; reconstituted reagent: -20°C for 4 weeks or 2–8°C for 2 days in the dark |
| Shelf Life | 12 months for lyophilized reagent at -20°C |
| Package Specifications | 10 mL (100 assays in 96-well), 100 mL (1,000 assays); also available in 5 mL and 50 mL vial sizes |
| Product Form | Lyophilized cake requiring reconstitution with provided buffer |
| Quality Control | Each lot standardized against purified recombinant caspase-3; luminescence linearity confirmed R2 ≥0.99 across the analytical range; Z-factor ≥0.7 for screening quality assessment |
| Key Features | Simple add-mix-read homogeneous protocol; non-lytic formulation preserves cell morphology for concurrent imaging; glow-type luminescence with >2 hour signal half-life; Ultra-Glo luciferase engineered for high thermal and chemical stability |
For research use only, not for clinical use.
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