ReadiUse™ MTS Reagent *5 mM aqueous solution*

1. Prepare the cells in a clear-bottom, 96-well plate.

2. Treat cells as desired.

3. Add 20 µL of the ReadiUse™ MTS Reagent working solution to each well.

4. Incubate at 37°C for 1 - 4 hours.

5. Monitor absorbance at OD = 490 nm.

For guidelines on cell sample preparation, please visit:

https://www.aatbio.com/resources/guides/cell-samplepreparation.html

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles

1. To prepare the ReadiUse™ MTS Reagent working solution, add PMS solution (not provided) to the ReadiUse™ MTS Reagent to achieve a final PMS concentration between 200 µM and 1000 µM. The optimal final concentration of PMS may be adjusted based on experimental requirements.
   
   Note: Protect the working solution from light by covering it with foil or storing it in a dark place.
   
   Note: For best results, this solution should be used within a few hours of its preparation.

1. Plate 5000 to 10,000 cells/well in a tissue culture microplate with a clear bottom. The suggested total volume is 100 µL for a 96-well plate or 50 µL for a 384-well plate.

2. Add test compounds to the cells and incubate for the desired duration (e.g., 24, 48, or 96 hours) at 37°C in a humidified incubator with 5% CO₂. For blank wells (medium without cells), add the same volume of test compounds. The recommended total volume is 200 µL per well for a 96-well plate or 100 µL per well for a 384-well plate.

   Note: Optimal cell density should be determined individually for each cell line. For proliferation assays, seed fewer cells; for cytotoxicity assays, start with a higher number of cells.

3. Add 20 µL of ReadiUse™ MTS Reagent working solution to each well of a 96-well plate, or 10 µL per well for a 384-well plate.

4. Incubate the plate at 37°C for 1 to 4 hours, protected from light.

   Note: Depending on the cell type and concentration, the optimal incubation time may range from 30 minutes to overnight. Adjust incubation conditions as needed for your specific experimental setup.

5. Monitor the absorbance increase with an absorbance plate reader at OD = 490 nm.

1. Seed cells in a clear-bottom tissue culture microplate. A total volume of 100 µL per well is recommended for a 96-well plate, or 50 µL per well for a 384-well plate.
   
   Note: Serially diluted suspensions of HeLa and Jurkat cells were prepared in a clear-bottom 96-well plate for the assay.

2. Add 10 µL of ReadiUse™ MTS Reagent working solution to each well of a 96-well plate, or 5 µL per well for a 384-well plate.

3. Incubate the plate at 37°C for 1 to 4 hours, protected from light.

   Note: Depending on the cell type and concentration, the optimal incubation time may range from 30 minutes to overnight. Adjust incubation conditions as needed for your specific experimental setup.

4. Monitor the absorbance increase with an absorbance plate reader at OD = 490 nm.