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| Product Name | Fluor 488 Caspase-3 Assay Kit for Live Cells |
| Catalog No. | RESK-0163 |
| Description | This live-cell assay kit contains the Fluor 488 Caspase-3 Substrate and the Ac-DEVD-CHO Caspase-3/7 Inhibitor. The Fluor 488 Caspase-3 Substrate provides an effective tool for detecting apoptosis based on caspase-3/7 activity and is suitable for fluorescence microscopy and flow cytometry analysis. In contrast to other FLICA-based fluorescent caspase substrates or fluorescent inhibitors, the supplied substrate detects caspase-3/7 activity without inhibiting the apoptotic process of intact cells. The substrate consists of a fluorescent DNA dye coupled to the caspase-3/7 DEVD recognition sequence. It is initially non-fluorescent and crosses the cell membrane into the cytoplasm. In apoptotic cells, caspase-3/7 cleaves the substrate to release a high-affinity DNA dye that migrates to the nucleus, labels DNA and emits bright green fluorescence. The substrate is bifunctional: it detects caspase-3/7 activity while visualizing the morphological changes of the nucleus during apoptosis. Fluor 488 staining is compatible with formaldehyde fixation and subsequent immunostaining. Spectral properties: Ex/Em = 500/530 nm (bound to DNA). |
| Intended Use | Detection of caspase-3/7 activity in living cells for apoptosis analysis by fluorescence microscopy or flow cytometry. |
| Principle / Technology | The substrate is a fluorescent DNA dye coupled to the caspase-3/7 DEVD recognition sequence. Initially non-fluorescent, it permeates the cell membrane and enters the cytoplasm. In apoptotic cells, active caspase-3/7 cleaves the substrate, releasing a high-affinity DNA dye that migrates into the nucleus, binds DNA and emits bright green fluorescence. Unlike FLICA-based fluorescent caspase substrates/inhibitors, the substrate detects caspase-3/7 activity without blocking apoptosis in intact cells. The substrate is bifunctional, reporting caspase-3/7 activity and simultaneously visualizing nuclear morphological changes during apoptosis. |
| Detection Method | Fluorescence detection by fluorescence microscopy or flow cytometry; Ex/Em = 500/530 nm (bound to DNA). |
| Sample Type | Live cells (live-cell staining only; not applicable to tissue sections). |
| Performance Range / Specifications | Visualizes morphological changes of the nucleus during apoptosis; staining is compatible with formaldehyde fixation and subsequent immunostaining experiments. |
| Specificity | The substrate carries the DEVD recognition sequence and is specifically cleaved by active caspase-3/7. |
| Reaction Conditions / Protocol | Apply the substrate to living cells and analyze by fluorescence microscopy or flow cytometry. Co-staining with Hoechst 33342 live-cell dye is optional and is mainly used to facilitate observation of the positive rate; choose according to experimental needs. Stained samples may be fixed with formaldehyde and subjected to subsequent immunostaining. |
| Components / Formulation | Fluor 488 Caspase-3 Substrate; Ac-DEVD-CHO Caspase-3/7 Inhibitor. |
| Storage Conditions | Store at 4 °C. |
| Shelf Life | At least 12 months |
| Package Specifications | 25 T 100 T |
| Shipping Conditions | Shipped on wet ice. |
| Expiration Date / Stability | Stable for at least 12 months when stored at 4 °C. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Suitable for fluorescence microscopy and flow cytometry; compatible with formaldehyde fixation and subsequent immunostaining; optional co-staining with Hoechst 33342. |
| Application Notes / Precautions | The kit is intended for live-cell staining only and cannot be used for staining tissue sections. Co-staining with Hoechst 33342 live-cell dye is optional; it facilitates observation of the positive rate and can be included according to experimental needs. The measured value represents caspase-3/7 activity, not concentration. |
| Recognition Site | DEVD (caspase-3/7 recognition sequence) |
For research use only, not for clinical use.
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