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The Janus family of tyrosine kinases (JAK) includes JAK1, JAK2, JAK3 and Tyk2. This family associates with a variety of cytokine receptors and is activated by ligand binding. Upon ligand binding to cytokine receptors, JAKs phosphorylate themselves and their associated receptors. This provides multiple binding sites for signaling proteins containing SH2 or other phospho-tyrosine-binding domains, including STATs, Shc, insulin receptor substrates and FAK. Thus, JAK kinases provide important links between cytokine/hormone receptors and downstream effector proteins, and efficient study methods are in high demand.
Figure 1: Measurement of phosphorylated and total JAK1.
3T3-L1 cells were cultured in 96-well plates for 48 hours and converted to adipocytes. Cells were then serum-starved for 16 hours, treated with 20 ng/ml LIF for 10 minutes and fixed. Total and phospho JAK1 were each assayed in triplicate using the phospho and total JAK1 antibodies included in the FACE JAK1 Kit. Data was plotted after correction for cell number (performed through use of Crystal Violet).
Antibody Specificities
The phospho-JAK1 antibody is specific for phosphorylated JAK1 and was raised against a synthetic phospho-peptide corresponding to residues surrounding Tyr1022 and Tyr1023 of human JAK1. This antibody recognizes JAK1 when phosphorylated at these sites and does not cross-react with related kinases. The total-JAK1 antibody recognizes JAK1 proteins regardless of the phosphorylation state.
