IGF1R Kit

Data was generated using TDA 2.0™-assembled IGF1R and a biotinylated peptide substrate derived from the native IGF1R substrate, IRS1. Enzyme and TDA 2.0™ at the indicated concentrations were incubated with 1 µM peptide substrate and 100 µM ATP for the times indicated. The HTRF signal was read 1 hour after the addition of detection reagents.

Using the TDA 2.0™ protein assembly reagent (violet bars), the activity of His-tagged IGF1R cytoplasmic fragment is improved significantly. Additionally, the TDA 2.0™ protein assembly reagent eliminates or reduces the requirement to use manganese chloride (biologically irrelevant salt) in many of these activity assays using soluble recombinant RTK fragments. In this production lot, specific activity of IGF1R measured at 100 nM is 69 U⁄mg (1 U = 1 nmol phosphate added per minute). With TDA 2.0™, the specific activity of IGF1R measured at 10 nM IGF1R is 1155 U⁄mg. Specific activity was calculated in the linear range of each experiment. For these assays, ATP was present at 100 µM, peptide substrate at 100 µM, and TDA 2.0™, where present, was 1⁄2500^th the enzyme concentration. Both assays were run at room temperature for the time indicated.