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Description
ATP is the primary energy currency in cells and serves as a reliable indicator of cellular metabolic activity. Its concentration correlates linearly with the number of viable cells, making ATP quantification an effective method for assessing cell viability or counting live cells.
This kit utilizes the ATP-dependent luciferase-catalyzed oxidation of luciferin to generate a chemiluminescent signal, enabling highly sensitive and quantitative measurement of intracellular ATP levels. Thanks to high-purity luciferin substrate, thermostable luciferase, and an optimized reaction formulation, this reagent exhibits enhanced cell lysis capability—making it particularly suitable for viability assessment in 3D microtissue cultures.
Upon addition to cultured cells or 3D spheroids, the reagent rapidly lyses cells, releasing ATP and triggering a stable luminescent reaction (as illustrated). The resulting light intensity is directly proportional to the amount of ATP—and thus to the number of viable cells—within a defined range, allowing accurate quantification of live cell numbers.
Features
- Convenient & Rapid: Ready-to-use detection reagent; stable signal and fast readout.
- High Sensitivity: Delivers superior signal-to-noise ratio, especially for 3D microtissue assays.
- Efficient Workflow: Data can be acquired within 30 minutes or less after reagent addition.
- Strong Lysis Capability: Optimized formulation ensures efficient disruption of 3D cell aggregates for complete ATP release.
- Scalable Throughput: Suitable for both low-sample experiments and high-throughput screening formats.
Components
|
Components No. |
Name |
40212ES10 |
40212ES60 |
40212ES80 |
|
40212 |
CellProlif-Glo™ 3D ATP Luminescent Cell Viability Assay Kit |
10 mL |
100 mL |
10×100 mL |
Storage
This product should be stored at -25℃~-15℃ for one year.
Figure
1. Application in 3D Spheroid Model
Figure 1. Cell viability analysis in 3D spheroid model using the CellProlif-Glo™ 3D ATP Luminescent Cell Viability Assay Kit.
HeLa cells (5,000 cells/well, 100 μL) were seeded in low-attachment 96-well plates and cultured for 4–6 days. After incubation, 100 μL of detection reagent was added, followed by shaking for 5 min and standing for 25 min. Then 100 μL of the reaction mixture was transferred to a white opaque 96-well plate for luminescence reading using a Nivo_Lum kinetic mode.
2. Application in Organoid Models.
Figure 2. Application of the CellProlif-Glo™ 3D ATP Luminescent Cell Viability Assay Kit in different organoid models.
The assay was successfully applied to ovarian cancer organoid model I, lung cancer organoid model II, and thyroid cancer organoid model II, demonstrating broad compatibility with various tumor organoid systems.
3. Strong stability
Figure 3. Stability evaluation of the CellProlif-Glo™ 3D ATP Luminescent Cell Viability Assay Kit.
The kit retained over 90% of its activity after storage at 4 °C for 60 days, and repeated freeze–thaw cycles (up to 10 times) showed negligible effect on product performance.
Documents:
Safety Data Sheet
Manuals
CCK-8 uses WST-8 [2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium] as a colorimetric indicator of cellular dehydrogenase activity. Living cells reduce WST-8 to a soluble orange formazan product — the amount of formazan generated is directly proportional to the number of metabolically active (viable) cells. ATP Luminescent kits measure intracellular ATP as a proxy for cell viability via bioluminescence.
Standard CCK-8 and 2D ATP assays are optimized for monolayer cultures. The 3D ATP Luminescent Cell Viability Assay Kit is specifically formulated for 3D spheroid models, incorporating enhanced lysis and solubilization chemistry to penetrate compact spheroid cores and recover total ATP signal. For standard CCK-8 with 3D cultures, extended incubation or matrix dissolution may be required — consult the datasheet.
Yes, provided the final DMSO concentration in the assay does not exceed 0.1–0.5% (v/v). Higher DMSO concentrations can independently affect cell metabolism and formazan/ATP generation, confounding viability readout. Always include DMSO vehicle controls at matched concentrations and confirm DMSO tolerance for your specific cell type before running compound screens.
CCK-8: Absorbance measurement at 450 nm (reference wavelength 620–650 nm). ATP Luminescent assays: Luminescence mode (no excitation filter). Ensure the instrument is equilibrated to room temperature and warmed up before measurement. For kinetic assays using CCK-8, the plate can remain in the incubator between reads — the assay is non-lytic and allows repeated measurement.
Yeasen Biotechnology offers flexible customization options for many of its assay kits and detection reagents, including custom lot sizes, bulk ordering, and application-specific formulation adjustments. Volume pricing, custom packaging, and kit bundling may be available depending on the product and intended workflow. A Certificate of Analysis (CoA) and lot-specific QC data are provided with every order. For inquiries regarding large-volume orders, custom configurations, or integration into automated workflows, please contact the BioHippo team for a tailored quotation.
