Description
Key facts
Detection method:Fluorescent,
Sample types:Tissue, Suspension cells, Microsomes, Adherent cells,
Assay type:Enzyme activity (quantitative),
Sensitivity:= 0.1 mU/well,
Assay Platform:Microplate
Product details:
UGT Activity Assay / Ligand Screening Kit (Fluorometric) (ab273331) enables rapid measurement of native or recombinant UGT activity in biological samples such as liver microsomes and can also be used to assess the effect of drugs and other novel compounds on UGT activity. The assay utilizes a highly fluorescent UGT substrate with a large Stokes shift (Ex/Em = 415/502 nm) that allows determination of UGT activity by tracking the drop in fluorescence emission as the substrate is converted into a non-fluorescent glucuronide. The multi-isozyme substrate is glucuronidated by virtually all of the pharmacologically-relevant mammalian UGT1A and UGT2B enzymes. UGT specific activity is calculated by comparing the fluorescence loss versus a control reaction performed in the absence of the required cofactor UDPGA. The kit includes the pore-forming peptide antibiotic Alamethicin, which allows the UGT Substrate and UDPGA to rapidly diffuse across lipid membranes to access the UGT active site located in the lumen of microsomes. For verification of modulation of UGT activity by test ligands, diclofenac, a competitive inhibitor of most human and rodent UGT isozymes, is also included. The assay is highly sensitive, simple to perform and high-throughput adaptable. This assay can detect less than 0.1 mU UGT activity in biological samples. The kit contains a complete set of reagents sufficient for performing 100 reactions at a 100 μl reaction volume.
Note: The UGT Substrate is glucuronidated by the majority of characterized human hepatic and non-hepatic UGT isozymes, aside from UGT1A4. Thus, the calculated UGT activity represents a composite of all of the isozymes expressed in a particular sample. In human liver microsomes, the major isozymes include UGT1A1, 1A3, 1A6, 1A9 and 2B7.
This product is manufactured by BioVision, an Abcam company and was previously called K692 UGT Activity Assay / Ligand Screening Kit (Fluorometric). K692-100 is the same size as the 100 test size of ab273331.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
The protein UGT also known as UDP-glucuronosyltransferase plays a significant role in the process of glucuronidation. Mechanically UGT facilitates the transfer of glucuronic acid from UDP-glucuronic acid to various substrates including hormones and drugs which increases their solubility in water. The mass of UGT proteins can vary depending on isoform types generally ranging from 50 to 60 kDa. UGT is mainly expressed in the liver but it is also found in the intestine kidneys and other tissues reflecting its comprehensive role in metabolism.
Biological function summary
The UGT enzymes function in detoxification processes by converting lipophilic substances into more water-soluble metabolites which can then be excreted from the body. UGT forms part of a multienzyme complex involved in phase II drug metabolism. It also interacts with other enzymes involved in conjugation pathways like sulfotransferases to ensure efficient and effective elimination of potentially harmful compounds.
Pathways
UGT enzymes play a pivotal role in the glucuronidation pathway which is a part of phase II of drug metabolism. This pathway connects to larger detoxification networks involving cytochrome P450 enzymes such as CYP3A4 which provide substrates for glucuronidation. Together these enzymes participate in xenobiotic metabolism impacting drug efficacy and safety by modifying drugs and endogenous compounds.
UGT enzymes have implications in inherited metabolic disorders like Crigler-Najjar syndrome which results from mutations in the gene encoding UGT1A1 leading to impaired bilirubin conjugation. UGT's activity is also linked to the development of hyperbilirubinemia or Gilbert's syndrome a milder condition characterized by intermittent jaundice. Additionally diseases such as cancer and the protein's interaction with other metabolic enzymes like GST (glutathione S-transferase) can modulate drug resistance affecting treatment outcomes.
