Application

• For high-throughput screening of FTase inhibitors and evaluation of drug modulators.

Key Features

• Safe. Non-radioactive assay.
• Homogeneous and convenient. “Mix-incubate-measure” type assay. No wash and reagent transfer steps are involved.
• High-throughput. A Z’-factor of 0.8 and higher is routinely observed in a 384-well format. Can be readily automated to assay thousands of samples per day.

Method

• FL340/550 nm

Samples

• Test compounds

Species

• All

Procedures

• 1 hr

Size

• 400 tests in 384well plate

Detection Limit

• NA

Shelf Life

• 6 months

More Details

• FARNESYLTRANSFERASE (FTase, EC 2.5.1.58) catalyzes the transfer of a farnesyl group from farnesyl pyrophosphate to the cysteine residue of the C-terminus of target proteins. When not properly regulated, farnesylated proteins, including the Ras superfamily of small GTPases, can lead to developmental disorders and cancer. Simple, direct and high-throughput inhibitor screening assays find wide applications for oncology research. BioAssay Systems’ EIFT-400 assay kit provides a convenient fluorimetric method to screen for potential FTase inhibitors. In this assay, FTase reacts with farnesyl pyrophosphate and a dansyl-peptide substrate with measurable fluorescence at λem/ex = 340/550 nm. Inhibition is determined by the decrease in fluorescence.

How can I set up the positive control for this assay?

We prepared a serial dilution of the 10mM Lonafarnib stock starting at 50 µM. Please see the protocol for an example graph of Lonafarnib inhibition.

We are not observing full inhibition in our compounds after the 10 minute incubation time. Is it ok to incubate for a longer period of time?

Yes, the initial incubation can be prolonged if needed.

Does your kit include FTase enzyme?

Our kit does not include enzyme. If the customer needs to provide their enzyme of choice for this assay. Recombinant rat FTase enzyme may be purchased from e.g. Jena Bioscience, Cat #PR-102.

For more detailed product information and questions, please feel free to email us or call us at 1-510-782-9988 x 2. For more general information regarding our assays, please refer to General Questions and Troubleshooting.

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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to request assay service.