What is the correct protocol for determining IC50 and CC50 of a drug?

04 September 2024 0 1K Report

I am planning to use Cycloheximide (CHX) chase assay to determine protein stability in myoblast cell line. I am going to perform this experiment for the first time so I am really confused about some points.

1. How much time is required for incubation with CHX before identifying IC50 and CC50. Is there any protocol to determine the optimized incubation time?

2. Do we need to determine the protein concentration for IC50 calculations? There are multiple youtube videos with IC50 calculations. They indicate that data is collected from spectrophotometry analysis after MTT/MTS assay but thats quite confusing for me. Please help me out if i am missing some point here.

3. Do I need to determine CC50 before IC50? Or both can be calculated simultaneously?

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