ATX inhibition is determined in the ATX inhibitor screening kit (cat #’s K-4200 or K-4200HTS). These assays utilize the fluorogenic substrate, FS-3, an LPC analogue, that is conjugated with both a fluorophore and a quencher. When ATX cleaves FS-3, the fluorophore becomes liberated from the quencher, resulting in increased fluorescence.

In this assay service, two screens are ran for each compound. In the initial screen, recombinant ATX is incubated with your inhibitor(s) or a 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 ATX inhibitors.

Test results will be provided as % inhibition or IC50. All data points will be run in duplicate and reported as an average. 

Sample Volume: 60uL/compound minimum is required at 40X the concentration to be tested. We can bring the compounds up in DMSO for an additional fee.

Sample Notes:

• The assay is sensitive to organic solvents. Limit to <2%.
• The assay is resistant to modest concentrations of DMSO. See Figure 1 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.

Storage and shipment of samples:

• Freeze all samples before shipping and ship on 3-5 lbs of dry ice, if possible.
• If samples cannot be frozen, use leakproof caps and vials.
• Label vials well. If vials leak, the solution can remove poor labeling. DMSO can erase or smear writing on labels.
• Contact your service coordinator and let them know the samples are on their way.

The minimum sample number for this service is 12. Example pricing is shown below and custom sample numbers can be accommodated. 

12 SAMPLES    $1,500.00
20 SAMPLES    $2,250.00

Standard pricing is for each sample to be run in duplicate wells. This means that 20 samples = 40 assay wells. If you need to run greater or fewer samples, contact us for pricing. If you want your samples run in triplicate, let us know! We will adjust pricing.