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| Product Name | RealTime-Glo Annexin V Apoptosis and Necrosis Multiplex Assay |
| Catalog No. | CATR-HMM-0051 |
| Description | A homogeneous live-cell assay combining two detection technologies to simultaneously monitor phosphatidylserine externalization and loss of membrane integrity in real time. The assay pairs an Annexin V fusion with a NanoLuc luciferase subunit for apoptosis detection with a cell-impermeant DNA-binding dye for necrosis. Both signals are measured from the same well without washing, fixation, or cell lysis. |
| Intended Use | Kinetic monitoring of apoptosis and secondary necrosis in the same cell population over time using a single addition, no-wash protocol. |
| Principle / Technology | Bioluminescence complementation for Annexin V binding detection plus DNA intercalating dye fluorescence for necrosis |
| Detection Method | Luminescence (NanoLuc, 460 nm) and fluorescence (DNA dye, 620 nm) |
| Sample Type | Adherent or suspension mammalian cells cultured in 96- or 384-well plates |
| Performance Range / Specifications | Apoptosis detection within 30 min of phosphatidylserine exposure; continuous monitoring up to 72 hr |
| Sensitivity / LOD | Detection of apoptosis induction by as little as 10 nM staurosporine in HeLa cells |
| Specificity | Annexin V fusion protein binds phosphatidylserine in calcium-dependent manner; DNA dye excluded from live cells with intact membranes |
| Reaction Conditions / Protocol | Add detection reagent directly to culture wells; mix briefly; measure luminescence (apoptosis) and fluorescence (necrosis) signals at desired intervals |
| Components / Formulation | Annexin V-LgBiT/SmBiT complementation reagent, cell-impermeant fluorescent DNA dye, calcium chloride, assay buffer |
| Storage Conditions | –20 °C for detection reagent; protect from light |
| Shelf Life | 6 months at –20 °C |
| Package Specifications | 100 assays (96-well) or 400 assays (384-well) |
| Product Form | Liquid reagents |
| Quality Control | Validated with staurosporine and anti-Fas antibody apoptosis induction; signal-to-background ratio ≥10 at 4 hr |
| Key Features | True real-time kinetic capability captures transient apoptosis events that endpoint assays miss |
| Purity | Reagent grade; dye purity ≥95% by HPLC; sterile where applicable |
| Concentration | As specified on product label; working concentrations optimized per protocol |
| Activity / Unit Definition | Signal-to-noise ratio and linearity verified on reference cell lines per lot |
| Molecular Weight | Varies by dye or reagent component as specified |
| Source / Origin | Synthetic dyes and tetrazolium compounds; recombinant enzymes where applicable |
| pH Range / Optimal pH | pH 7.2–7.4 for cell-based assays |
| Shipping Conditions | Cold packs; protect light-sensitive reagents from light |
| Expiration Date / Stability | 6–12 months at recommended storage temperature |
| Regulatory / Compliance | Research use; manufactured under ISO 9001 |
| Compatibility | Compatible with adherent and suspension mammalian cell culture in 96-well and 384-well formats |
| Recommended Buffer System | PBS or phenol red-free complete medium as specified in protocol |
| Application Notes / Precautions | Optimize cell seeding density; include appropriate vehicle controls; avoid phenol red interference with absorbance-based assays |
| Batch-to-Batch Consistency | Signal linearity R² ≥0.99 with reference cell number standard curve per lot |
For research use only, not for clinical use.
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