Autotaxin (ATX) Inhibitor Screening Service

 The Autotaxin (ATX) Inhibitor Screening Service utilizes the ATX Inhibitor Screening kits (cat #’s K-4200 or K-4200HTS) to determine if your compound inhibits ATX. ATX is an extracellular enzyme that cleaves lysophosphatidylcholine (LPC) to generate the phospholipid growth factor lysophophatidic acid (LPA). LPA is involved in a variety of biological functions, such as angiogenesis, wound healing, vascular and nervous system development, reproduction, pain transmission, immunological competence and the regulation of blood pressure. Altered LPA biology is associated with a number of diseases. Likewise, autotaxin has been implicated in a number of disorders, including a variety of cancers, obesity, arthritis, multiple sclerosis, Alzheimer’s disease and neuropathic pain.

• Echelon has 25 years of assay development expertise and 15 years experience in the ATX field.
• Your compounds are cataloged and stored according to your requirements.
• The ATX inhibitor Screening Service will run each compound in duplicate, in two screens.
• Results are provided as % inhibition or IC50, if a full inhibitor curve is ran.
• Usually, results are provided 2 weeks from receipt of samples. We can accommodate tighter schedules if needed.

Contact us for a quote on your custom assay service

Description

Assay Details

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 Considerations

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.

Cost

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.

Contact Us

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Publications

Citations listed are from the Autotaxin Inhibitor Screening Kit (catalog number K-4200).

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