CyQuantiFluorTM Cell Viability Detection Kit

Youai Bio's CyQuantiFluor™ Cell Viability Assay Kit includes a DNA‑specific binding dye and a background‑quenching dye. The DNA‑specific binding dye exhibits only weak fluorescence in the absence of DNA; however, once it crosses the live cell membrane into the cell and specifically binds to intracellular DNA—primarily within the nucleus—it emits much stronger fluorescence. The fluorescence intensity is proportional to the amount of bound DNA, allowing direct quantification of live cells. The background‑quenching dye cannot penetrate the live cell membrane; instead, it quenches fluorescence arising from DNA bound by the dye but originating outside live cells, such as DNA released from dead cells with ruptured membranes, thereby ensuring that only fluorescence from the dye bound to DNA within live cells is detected. The CyQuantiFluor™ Cell Viability Assay measures the DNA content of cells with intact membranes directly, independent of cellular metabolic status, enabling more accurate counting of live cells. Moreover, its specific binding to DNA further enhances the precision of the results.

The CyQuantiFluor™ Cell Viability Assay Kit is a homogeneous reagent that requires only a single addition for detection, thereby reducing experimental errors that may arise from multiple additions. This reagent features a high signal-to-noise ratio, excellent reproducibility, and superior stability. It can quantify the number of viable cells in both cell proliferation and cytotoxicity assays, making it particularly well-suited for tumor cell proliferation studies and high-throughput screening of compounds.

CyQuantiFluor^TM The components and specifications of the cell viability assay kit are as follows. Among them,for the96-well plate and the384-well plate, the number of detection reactions is calculated based on a medium volume of100 μLper well and, with the test reagent added at100 μLand25 μLper well, respectively.

Specifications	Components and Specifications	Assay96-Number of Wells in 96-Well Plate	Assay384-Number of Wells in 384-Well Plate
250 assays	DNAbinding dye:0.1 mL background-quenching dye:0.5 mL	250	1,000
1,250 assays	DNAbinding dye:0.5 mL background-quenching dye:2.5 mL	1,250	5,000
12,500 assays	DNAbinding dye:5 mL background-quenching dye:25 mL	12,500	50,000

Applicable under most experimental conditionsCyQuantiFluor^TMIt is not recommended to directly compare the absolute readings of samples using the cell viability assay kit. It is advised to set the same reference on each experimental plate—for example, in tumor cell proliferation assays, use the same toxic reference compound and solvent carrier control. Normalize the sample readings against those of the toxic reference compound and solvent carrier control before data processing and comparison.

The following uses as an example96well cell culture plates fordetection.

1)On a96-A transparent black-bottomed 96-well cell culture plate is seeded with experimental cells at an appropriate density.100μL

2)Perform cell treatment according to experimental requirements; for example,in tumor cell proliferation experiments, add the corresponding test compound and incubate for a certain period.

3)After cell treatment,take outCyQuantiFluor^TMcell viability assay kitand performcell viabilitydetection

4)2x CyQuantiFluor^TMPrepare the detection reagent: you can useHBSS (Hank's buffered saline solution), PBS,Preparation of culture media and other solutions, with or without serum2xTest reagent. Dispense the above solution as needed, then add250xof/'sDNACombining dyes and50xYou can simply use a background‑blocking dye, for example, to test a piece.96-The breadboard can be provided as needed.Preparation12 mLof/'s2xTesting reagents can be directed to11.7 mLof/'sHank's Balanced Salt SolutionAdd toforty-eightmuLof/'sDNACombination dyeand240muLof/'sBackground-suppressing dyeThat's all. Shake to mix.

5)To each well containing100muLThe culture medium's96-ConfuciusAdd to the experimental cell plate100muL 2xDetection reagents. If each well contains a different volume of culture medium, it should be maintained…The volume ratio of culture medium to added reagent per well is1 : 1

6)37℃ 5% CO₂Incubation in an incubator. The test reagent can be added.0.5 hour - 6 hoursPlate reading can generally be done in1 hour. The optimal reading time depends on the specific cells and experimental conditions and can be optimized through preliminary experiments. Fluorescence signal intensity typically reaches its peak within30–60 minutesand remains essentially stable for up to6 hours. Note that if the test cells are suspended, ensure they have settled at the bottom of the assay plate before reading; if necessary, gently centrifuge the plate to bring the suspended cells to the bottom.

7)Use aFITCfilter(Em 490nm/Ex 520nm)or a similar filter to read the fluorescence signal from thebottom

1) Mixing different batches is not recommended.
2) Without rigorous validation, it is not recommended to alter the volume of the test reagent.
3) For research use only.