Protocol SDS

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

• Compounds that affect farnesyltransferase activity

Species

• All

Procedures

• 60 min

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.