The Echelon Autotaxin (ATX) Inhibitor Assay Service utilizes one of Echelon’s Autotaxin Inhibitor Screening kits depending on the scale of the project (catalog#’s K-4200 or K-4200HTS) . These kits use the fluorogenic ATX substrate, FS-3. FS-3 is an LPC analogue that is conjugated with both a fluorophore (fluorescein) and a quencher. In its native state the quencher interferes with fluorescein’s fluorescence. Once ATX cleaves FS-3, fluorescein becomes liberated from the quencher, resulting in increased fluorescence.
In this assay service, two screens are run for each sample. In the initial screen, recombinant Autotaxin is incubated with your inhibitor(s) and an internal control inhibitor (BrP-LPA). The substrate, FS-3, is then added subsequently and fluorescence is measured over time. These assays are continuous (not end point) and have the advantage of being less susceptible to error arising from mistiming in the addition of reagents. The secondary screen is performed to evaluate whether a sample hit, revealed in the initial screen, is a false positive due to interference with the assay. The sample inhibitors that are positive in the initial screen and negative in the secondary screen are potential Autotaxin inhibitors.
Test results will be provided as % inhibition or IC50. All data points will be run in duplicate and reported as an average. For more information on the Autotaxin Inhibitor Screening kit, please see Technical Data Sheet (TDS) for catalog # K-4200 or K-4200HTS.
-The assay is sensitive to organic solvents. Limit to <2%.
-The assay is resistant to modest concentrations of DMSO. See figure 2 on the K-4200 Technical data sheet.
-We will run controls for all solvents used in your sample panel. This may add additional costs.
-It may be preferred to send us vials of your sample solvent.
Sample Amount: 60uL/sample minimum is required.
Samples need to be in solution at the concentrations to be tested.