3D Cell Viability Assay Kit (ATP Luminescence) servicebio G1615-10ML
$50.00
In stock
SKU
G1615-10ML
Product information
| product name | Identification of product |
| model | 3D Cell Viability Assay Kit (ATP Luminescence) |
| G1615-100T | 10 mL |
Descirption/Introduction
The 3D Cell Viability Assay Kit (ATP Luminescence) is a cell viability detection reagent based on a luciferase system. The detection reagent has been optimized for stronger lysis capability, effectively penetrating into the interior of spheroids formed by 3D cultures, detecting the viability of microtissues produced by 3D cell cultures. When the product is added to cell cultures, it causes the cell clusters to lyse and release ATP, thereby emitting a stable "glow-type" luminescent signal. The intensity of this signal is directly proportional to the amount of ATP within a certain range, indirectly reflecting the number of living cells in the sample. Therefore, this product can be used for quantitatively detecting the number of living cells.
This product is optimized for the viability testing of 3D cell spheres obtained from various cultures, such as cell suspension plates, ultra-low adsorption plates, matrix gel-coated and agarose-coated 3D cell spheres. It is a ready-to-use solution, which can be used immediately without any preparation, after adding cells, it can be simply incubated and tested, which is very suitable for high-throughput screening of large-scale samples.
Storage and Transportation
Shipping with dry ice; Store the kit at –80°C for 12 months.
Component
| Component Number | Component |
| G1615-100T | G1615 |
| 3D Cell Viability Assay Reagent | 10 mL |
| Manual | 1 pc |
Assay Protocol / Procedures
1. Cell viability/proliferation/toxicity assay
2. Cell preparation:Inoculate a certain amount of cell suspension 100 µL in 96-well plate (if using 384-well plate, inoculate 25 μL cells per well), determine the amount of cell inoculation and inoculation time according to the culture method, the number of days of cultivation, the size of the 3D cell spheroplasts needed, etc. If necessary, add drugs to treat the cells.
3. Reagent Preparation: After dissolving the reagents, aliquot them appropriately to avoid repeated freeze-thaw cycles. Bring an appropriate amount of reagent to room temperature before use. Gently invert up and down to mix and obtain a homogeneous solution.
4. During detection, equilibrate the cell culture plate to room temperature (approximately 10 minutes). Directly add the reagent in equal volume to the cell culture medium (e.g., for a 96-well plate containing 100 μL of culture medium per well, add 100 μL of ATP luminescence 3D cell viability assay reagent; for a 384-well plate, 25 μL per well). Shake at room temperature for 5 minutes to promote cell lysis.
5. Incubate at room temperature (approximately 25°C) for 25 minutes to stabilize the luminescent signal.
6. Detect chemiluminescence using a multifunctional microplate reader with chemiluminescence detection capabilities. Set the corresponding parameters according to the instrument requirements; generally, the detection time is 0.25-1 second or longer per well, specifically adjusted based on the instrument's sensitivity.
7. Calculate the relative cell viability directly from the chemiluminescence readings, or calculate the amount of ATP through an ATP standard curve to determine the relative cell viability.
8. Vitality Calculator:
Cell viability (%) = [Aexperiment-Ablank]/[ Acontrol-Ablank]×100
Inhibition rate (%)=[Acontrol-Aexperiment]/[ Acontrol-Ablank]×100
A experimental:Treated cells + 3D cell viability assay reagent
A blank: Cell culture medium (no cells) + 3D Cell Viability Assay Reagent
A control: Normal untreated cells + 3D cell viability assay reagent
Note
1. Repeated freeze-thaw cycles will reduce the detection efficiency. For optimal use, after the first thaw, aliquot and store appropriately, ensuring that the containers are free from ATP contamination. Long-term storage should be at -80℃..
2. The activity of luciferase is quite sensitive to temperature; therefore, both cells and detection reagents need to be equilibrated to room temperature before the assay. Detection reagents can be dissolved and mixed in a room temperature not exceeding 25℃ or in a water bath at 20-25℃ before use.
3. Use either white or black 96-well plates or 384-well plates for testing. Using regular transparent 96-well or 384-well plates may cause interference between adjacent wells. Alternatively, 3D cultures can be grown in ultra-low attachment dishes or multi-well plates designed for 3D culture, then transferred to ordinary white or black multi-well plates for testing.
4. High concentrations of drug solvents may interfere with the luciferase reaction, affecting the chemiluminescence signal. This can be mitigated by including control wells containing the solvent in the cell culture medium. Testing has shown that DMSO content within 2% of the final reaction volume does not affect the reaction.
5. If cells are cultured for an extended period, evaporation must be considered. To address this, avoid using the outermost wells of the 96-well plate as they are most susceptible to evaporation; instead, fill them with an equal amount of PBS, water, or culture medium. Additionally, placing the 96-well plate closer to the water source inside the incubator can help mitigate evaporation.
6. Please wear a lab coat and disposable gloves when operating.
This product is for research only, not for clinical diagnosis!
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