Transcreener® UDP FP Assay
BellBrook Labs Transcreener UDP FP Assay
The BellBrook Labs Transcreener® UDP FP Assay is a competitive fluorescence polarization (FP) assay based on the detection of UDP and therefore is compatible with any enzyme class that produces UDP, including glucosyltransferase, galactosyltransferase, glucuronyltransferase, N-acetylglucosamyltransferase, N-acetylgalactosyltransferase, xylosyltransferases, and glycogen, cellulose, lactose and hyaluronan synthases. The Bell Brook Lab's Transcreener UDP FP Assay is a simple one step homogenous detection assay, and is extremely flexible with regard to substrate concentration (1 to 250 μM).
• Universal. One set of reagents for any UGT isoform
• Efficient. Thousands of compounds can be evaluated in one day.
• Easy. Homogenous “mix and read” assay format.
• Cost effective. Low volume assays means minimal cost associated with reagents.
• Robust. Z' factor values routinely over 0.7.
There are over 200 glycosyltransferases encoded in the human genome, and most use uridine diphosphate (UDP) -activated sugars as the donor. From a functional standpoint, the reactions they catalyze can be divided into three major types: biosynthesis of disaccharides or polymers such as starch or hyaluronan, posttranslational modification of proteins, and glucuronidation of small molecules, including endogenous hormones and xenobiotics.
All three types of glycosyltransferases are of interest in drug discovery. The role of the approximately 15 hepatic UDP-glucuronosyltransferases (UGTs) in drug metabolism has been an area of focus for several years. More recently, protein glycosyltransferases and those involved in synthesis of biopolymers have come under focus as therapeutic targets, for cancer and lysosomal storage diseases. Bacterial glycosyltransferases involved in cell wall synthesis are also being targeted for development of new anti-infectives.
In the Transcreener UDP FP Assay, UDP produced in the enzyme reaction is detected using a competitive fluorescence polarization immunoassay. Enzyme activity is indicated by a decrease in fluorescence polarization values of reactions containing acceptor-substrate relative to negative control reactions lacking an acceptor.
The Transcreener UDP FP Assay has been designed to provide Z'-factor values routinely greater than 0.7 in 384-well, 20 µL reaction format using hyodeoxycholic acid as acceptor-substrate and UGT2B7. This assay has demonstrated great flexibility for UGT source, and donor- and acceptor-substrate concentrations.
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