In cells

Use at concentration up to 5 µM. For in vitro studies a 10 mM stock solution in DMSO is used and mixed directly into culture medium containing 10% FBS or HS. The presence of serum facilitates the compound to stay in solution and the equilibrium between serum binding and permeation into the cell is sufficient to enable target engagement inside the cell. Note: For biochemical binding assays, i.e., TR-FRET use as described in PMID: 25313317. Due to sub-nM potency at these conditions solubility is not an issue, and the addition of serum is not required.

Use the recommended concentration to avoid off-target toxicity and/or off-target activity. Using concentrations that are higher than recommended could lead to false positive or confounding results.

In vivo

Use up to 5 mkd (IP) for probe. A protocol for the formulation of the probe is included in the additional probe information pdf that can be downloaded from the "Probe overview tab". Note: The control has not been characterized for in vivo use.
A-1155463 causes mechanism-based, rapid and reversible thrombocytopenia in mice. In vivo effects: a) Platelet reduction 6 h after a single IP dose of A-1155463 (5 mg/kg) in female tumor-free SCID-Beige mice; b) 50% tumor growth inhibition after 14 days of IP dosing (5mg/kg) in SCID mice bearing H146 tumors.