• This product exhibits exceptional stability in luminescent signals. The luminescent signal stability of this product is notably excellent, with an overall signal fluctuation typically not exceeding ±10% within 1 hour. In terms of luminescent stability, it outperforms similar products significantly, making it particularly suitable for high-volume, continuous cell viability measurements in 96-well or 384-well plates when a large number of samples need to be analyzed. The stability of the luminescent signal may vary slightly due to factors such as cell type, cell culture medium, external temperature conditions, and more. For instance, in adherent NIH3T3 and HeLa cells, signal intensity may initially increase and then decrease with prolonged reaction time, while in suspended Jurkat cells, the signal intensity may continuously decrease slightly with extended reaction time. However, the luminescent signal typically fluctuates within ±10% over 60 minutes and within ±25% over 120 minutes, with a signal half-life lasting up to 5 hours.
• This product offers high detection sensitivity and a wide linear range, providing excellent linearity within a range of 10 to 50,000 cells. When using this product to detect as few as 25 cells per well, the luminescent intensity can reach approximately twice that of the blank wells. The upper limit of detection quantity may vary significantly among different cell types. For some cells with exceptionally high ATP content, linearity may not be maintained beyond 50,000 cells, but the chemiluminescent readings will continue to increase.
• This product is simple to use, provides stable readings, and offers a fast detection process, typically requiring only about 10 minutes to complete. It is more straightforward and quicker compared to common methods for assessing cell viability such as MTT, AlamarBlue, Calcein-AM, WST-1, CCK-8, and CCK-F. To perform the cell viability assay with this product, simply dissolve the detection substrate (lyophilized powder) provided in the assay kit using the supplied assay buffer to create the Steady Luminescent Cell Viability Assay reagent. Then, mix this reagent with an equal volume of cultured cells. After a 10-minute incubation, you can proceed with chemiluminescent detection. There is no need to wash the cells, change or remove the culture medium. Additionally, the luminescent signal is highly stable, with an overall signal fluctuation typically not exceeding ±10% within 1 hour.
• This product offers excellent stability. It can be stored long-term at -20°C, and its stability remains strong even after preparation as the Steady Luminescent Cell Viability Assay reagent. Subjecting it to up to 5 freeze-thaw cycles has little to no impact on its detection performance, and even after 10 freeze-thaw cycles, the decrease in detection effectiveness is not more than 10%. When stored at 4°C for 3 days, the Steady Luminescent Cell Viability Assay reagent shows no significant impact on detection performance, and after 7 days, the decrease in effectiveness does not exceed 10%. Storing the reagent at room temperature for 1 day preserves over 80% of its detection performance, while at 37°C, it retains over 60% of its detection performance after 1 day. Even when retaining only about 50% of its detection effectiveness, the Steady Luminescent Cell Viability Assay reagent still meets the requirements for various routine assays.
• This product offers flexibility and convenience in its usage. It is not only suitable for the testing of small sample quantities but also highly suitable for high-throughput screening of a large number of samples.

Additional Notes

• The activity of the luciferase enzyme is sensitive to temperature, so both the cells and the assay reagent should be equilibrated to room temperature before conducting the reaction. You can thaw and mix the detection buffer in ambient temperature (not exceeding 25°C) or in a water bath set at 20-25°C before mixing it with the detection substrate to prepare the Steady chemiluminescence assay reagent.
• The Steady Luminescent Cell Viability Assay reagent in this kit contains luciferase, and repeated freeze-thaw cycles can gradually deactivate it. Although testing has shown that the Steady Luminescent Cell Viability Assay reagent remains unaffected by up to 5 freeze-thaw cycles, it is recommended to prepare and use it fresh for stability and optimal performance. Any unused assay reagent can be aliquoted and stored, but ensure that the aliquoting containers are not contaminated with ATP. During repeated freeze-thaw cycles, slight precipitation may occur in the assay reagent. In this case, bring it to room temperature and attempt to dissolve it as much as possible. If there are still insoluble particles, you can remove them by centrifugation, and testing has shown that this does not affect subsequent detection results.
• High solvent content in the test drug may interfere with the luciferase reaction and affect the chemiluminescent signal. You can exclude solvent interference by using cell culture medium containing the solvent as a control well in your assay. Testing has shown that a DMSO content of up to 2% in the final reaction system does not affect the reaction.
• For the detection, use white or black 96-well or 384-well plates suitable for cell culture. Using regular transparent 96-well or 384-well plates can lead to interference between adjacent wells. It is recommended to use 96-Well Black Opaque Plates.
• The user manual provides a method for detecting ATP standards. In practical cell viability testing, it is often unnecessary to measure ATP standards.
• This product is intended for scientific research by professionals only and should not be used for clinical diagnosis or treatment, food, or pharmaceuticals, and should not be stored in regular residential areas.
• For your safety and health, please wear laboratory attire and disposable gloves during operations.

Storage

Store at -20°C in the dark, and it remains effective for at least two years. The Steady Luminescent Cell Viability Assay reagent, prepared by mixing the detection buffer and substrate, retains its effectiveness for at least one year when stored at -20°C in the dark and for at least two years when stored at -80°C in the dark.

Only for research and not intended for treatment of humans or animals

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