1. Prepare a 12mM MTT stock solution by adding:
a. 1mL of sterile PBS to 5mg MTT
b. Mix by vortexing or sonication until dissolved
c. Once prepared, the MTT solution can be stores for 4 weeks at 4 degrees Celsius, protected from light
2. Prepare SDS_HCL solution
a. Add 10mL of 0.01M HCL to 1g of SDS.
b. Mix the solution gently by inversion or sonication until the SDS dissolves.
c. Once prepared, the solution should be used promptly.
1. Mix 1mL 12mM MTT with 10mL media
2. Aspirate old media from plate
3. Add 100ul per well of the MTT/Media solution to each well of a 96-well plate.
4. Incubate for 4 hours.
5. Add 100ul of the SDS-HCL solution to each well and mix thoroughly.
6. Incubate the plate for 4 hours. Longer incubation will decrease the sensitivity of the assay.
7. Mix each sample again using a pipette.
8. Use the Plate reader and read absorbance at 570nm.
a. Ensure the bulb is warmed up
b. Click on Re-read
c. Save and Export results to Excel
d. Title with Date, Initial and MTT.