From a compound to its IC50

What you will be able to do

• Calculate the mass of a salt form to weigh for a stock of given molarity and volume, and state the direction and size of the error made by weighing on the parent's molar mass.
• Plan a serial dilution that reproduces a published dose series, identify the step that breaks the bench pipetting rule, and choose a remedy that fixes it.
• Normalise each plate to its own controls, and interpret Z′ as a property of the controls, not of the compound's curve.
• Report a fitted relative IC50 with its interval, its context (cell line, readout, exposure, source) and the compound the source says was tested, and distinguish it from the absolute IC50 and from a binding constant.

The steps

1. 1. Molecule

   Tell the salt you weigh from the parent the result is about.

2. 2. Calc

   Make a stock of the salt at a stated molarity.

3. 3. Calc

   Plan the series the published screen used, and check it can be pipetted.

   Before this step: Dilutions and Buffers

4. 4. Assay

   Place the plan on the published plates and read their controls.

5. 5. Assay

   Fit the curve and read the relative IC50 with its interval.

   Before this step: Dose–Response

6. 6.

   See where the result is filed, and why.