Our tyrosine kinase assays are produced using a cell-based platform first developed by Mathey-Prevot and colleagues (learn more). The system relies on constitutively activated kinases to confer IL3-independent growth on the mouse Ba/F3 Pro B or FDC-P1 myeloid cell lines. Constitutive activation is achieved by fusing the kinase domain to donor proteins that typically (though not always) facilitate homomeric oligomerization and subsequent trans-phosphorylation. Blocking the activity of the activated kinase causes cell death, which we assess by measuring residual ATP.
Interestingly, kinase activities and their responses to inhibitors are influenced by the activating fusion protein and the cell type in which the kinase is expressed.