How to use chemical probes

How do I know what concentration I should use for my experiment?

The Chemical Probes Portal provides guidance on the maximum concentration that a probe should be used in cells.

However, when applying a chemical probe that was validated in one cellular system to another, it is important to consider several factors. Is the target expressed in the new system at levels comparable to those in the original? If not, the concentration of the probe that you will need to impact the activity of the target will differ.

Similarly, are likely off-target proteins expressed at the same levels? It is important to balance the amount of the probe that you use in cells with its selectivity, and the best concentration may need to be determined empirically by assessing on- and off-target activity profiles.

Similarly, we recommend validating that the chemical probe engages its intended target when moving to a new cellular system. Proteins can adopt different conformations and participate in distinct complexes in different cells; thus, it is essential to demonstrate that the protein target is accessible by the probe in the new system before proceeding with new experiments.