JAK1 (JH2 Pseudokinase Domain) Inhibitor Screening Assay Kit

The JAK1 (JH2 Pseudokinase Domain) Inhibitor Screening Assay Kit is a fluorescence polarization-based assay designed for screening and profiling small molecules that displace the fluorescently labeled probe (JH2 probe 1) from the JAK1 (Janus kinase 1) JH2 Pseudokinase Domain. This kit comes in a convenient 384-well format, with enough recombinant human JAK1 (JH2 Pseudokinase Domain), buffer, and fluorescently labeled JH2 Probe 1 for 384 reactions.Figure 1: Illustration of the assay principle.The assay is based on the competition between a test compound and the JH2 probe for the purified JAK1 (JH2 Pseudokinase Domain) . JH2 Probe 1 is incubated with JAK1 (JH2 Pseudokinase Domain) in the presence or absence of an inhibitor. When the JH2 Probe 1 is bound to the JH2 Pseudokinase Domain it has high FP due to its restricted movement. A competitive inhibitor prevents the probe from binding to the JH2 Pseudokinase Domain, therefore most of the probe is in free form and has low FP (fluorescence polarization) . Compared to a control without inhibitor, FP decreases proportionally to the inhibitor concentration.This assay requires a fluorescent microplate readercapable of measuring fluorescence polarization (FP) to read the FP signal. For more information FP technology, visit our Tech Note:FP, assay principles and applications.