PIM3 Cellular Phosphorylation Assay (intracellular kinase activity assay) for compound screening and profiling in intact cells
Pim kinases are constitutively active serine/threonine kinases that promote growth factor-independent proliferation by phosphorylation and subsequent inhibition of a range of cellular proteins. Activity of Pim kinases is physiologically regulated via protein stability. Especially Pim1 and Pim2 kinase have been found to be overexpressed or mutated in a variety of human tumors including B cell lymphomas, leukemias, prostate cancer and oral cancer.
PIM3
PIM3
HEK293T
Transfected
Transiently transfected Human embryonic kidney cells (HEK) express full length Pim3 kinase as well as Pim-substrate Bad labeled with a myc-tag. Constitutive activity of exogenous Pim3 results in phosphorylation of exogenous Bad at Ser112. Upon compound incubation, cells are lysed and Bad phosphorylation is quantified by Sandwich-ELISA technique. The assay is validated based on the cognate inhibitor of Pim kinases AZD1208 (see Fig.1).
Substrate phosphorylation as a readout of intracellular kinase activity via ELISA
Freiburg, Germany
More information can be found on our website Cellular Phosphorylation Assay Services.
Reference compound IC50 for PIM3
![Reference compound IC50 for PIM3](/wp-content/uploads/2023/11/PIM3_cell_phospho_freiburg.png?itok=Z-mhjp_c)
Cognate Pim inhibitor AZD1208 blocks Pim3 and inhibits phosphorylation of Bad at Ser112 with highly reproducible IC50 values. The graph shows a representative result.