MEK1/2 inhibitor treatment

If the blood was derived from a clinical trial where probands were treated with MEK1/2 inhibitors, this section can be skipped. In this case, continue immediately with the PMA stimulation.

Example: six groups: baseline control, positive control, solvent control, and three groups treated with different concentrations of the MEK1/2 inhibitor.

Note: Always prepare a baseline (no treatment, no stimulation), positive (no treatment, PMA stimulation), and solvent control (treatment with the solvent in which the MEK1/2 inhibitor is prepared, PMA stimulation), to assess minimal and maximal MEK1/2 activation, as well as any solvent effects.

Example: The desired treatment concentration with the MEK1/2 inhibitor is 10 μg/ml in 10 ml cell suspension, therefore prepare at least 100 µl of a 1 mg/ml WS in PBS.

Note: Prepare a solvent WS with the same solvent concentration as present in the MEK1/2 inhibitor WS.

Attention: In case of DMSO used as a solvent, do not expose the cells to concentrations exceeding 1% DMSO to avoid cytotoxic effects. Keep in mind that the subsequent stimulation with PMA is carried out in a final DMSO concentration of 0.1%.

Example: Add 100 µl WS to PBMCs suspended in 10 ml plasma.

Attention: Do not add anything to the baseline and positive control but mix gently.