UA-Glo® Luminescent Cell Viability 2.0 Assay

The Homogeneous Ready-to-Use Cell Viability Assay Kit 2.0 is used for the quantitative detection of intracellular ATP content, and the ATP content is proportional to the number of viable cells. After the first dissolution, the reagent can be stored at 4°C for two months, with the detection signal intensity reduced by <15% and no loss of functionality. This reagent features high signal-to-noise ratio, good repeatability, and excellent stability. The ready-to-use formula eliminates the experimental steps of first lysing and then detecting, thereby further reducing errors caused by frequent sample addition. Moreover, its stable glow signal makes this product particularly suitable for high-throughput cell proliferation detection and compound screening.

Luciferase, luciferin, and buffer are mixed and filled into brown bottles of 10 ml, 100 ml, or 500 ml. The specifications are as follows:

1. Cell Preparation
1) Seed the cells to be tested at an appropriate density in a 96-well or 384-well cell culture plate. It is recommended to use a white plate.
2) If the experiment is to determine the effect of compounds on cells, add the test compounds at appropriate concentrations to the wells of the cell plate. The concentration of organic solvents in the culture medium should be kept below 2%. Continue culturing for an appropriate time according to experimental needs.

2. Cell Viability Assay
1) Take out the Cell Viability Assay Reagent 2.0 [Note 1], equilibrate to room temperature (22°C-25°C) [Note 2], and mix gently by shaking.
2) Take out the cell culture plate to be tested and equilibrate to room temperature (22°C-25°C).
3) Add 50µL of Cell Viability Assay Reagent 2.0 to 100µL of cells in a 96-well plate, or 10µL of reagent to 20µL of cells in a 384-well plate [Note 3].
4) Shake the assay plate for 2 minutes to fully lyse the cells, and place it in the dark to equilibrate the reaction for 10 minutes.
5) Read the fluorescent signal on a fluorescence microplate reader [Note 4].

1) After the first use of Cell Viability Assay Reagent 2.0, it can be aliquoted and stored at -20°C or below in the dark. The signal intensity of the reagent decreases by <10% after 5 freeze-thaw cycles, and its function remains intact. After the reagent is dissolved for the first use, when left at room temperature (22°C) for 10 days or at 4°C for two months, the signal intensity decreases by <15%, and its function remains intact.
2) The luciferase reaction in Cell Viability Assay Reagent 2.0 is sensitive to temperature changes. The reagent and assay plates need to be equilibrated to room temperature (22°C-25°C), and the temperature should be kept constant (±1°C) during the test.
3) It is not recommended to arbitrarily change the amount of reaction reagents without strict verification. The volume ratio of cell culture medium to assay reagent should be 2:1.
4) Cell Viability Assay Reagent 2.0 has different signal decay rates for different cell types, with a signal half-life ranging from 1.5hr to 4hr. It is recommended that the plate reading time does not exceed 2hr.
5) This product is for research use only.