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.
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- Frontiers in pharmacology 14:11775412023Ontogeny of CYP3A and UGT activity in preterm piglets: a translational model for drug metabolism in preterm newborns.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLaura Buyssens,Allan Valenzuela,Sara Prims,Miriam Ayuso,Thomas Thymann,Chris Van Ginneken,Steven Van CruchtenPubMed 37124224
Key facts
- Detection method
Fluorescent
- Sample types
Tissue, Suspension cells, Microsomes, Adherent cells
- Assay type
Enzyme activity (quantitative)
- Sensitivity
= 0.1 mU/well
Target data
Function
Isoform 1UDP-glucuronosyltransferase (UGT) that catalyzes phase II biotransformation reactions in which lipophilic substrates are conjugated with glucuronic acid to increase the metabolite's water solubility, thereby facilitating excretion into either the urine or bile (PubMed:12181437, PubMed:15472229, PubMed:18004206, PubMed:18004212, PubMed:18719240, PubMed:19830808, PubMed:23288867). Essential for the elimination and detoxification of drugs, xenobiotics and endogenous compounds (PubMed:12181437, PubMed:18004206, PubMed:18004212). Catalyzes the glucuronidation of endogenous estrogen hormones such as estradiol, estrone and estriol (PubMed:15472229, PubMed:18719240, PubMed:23288867). Involved in the glucuronidation of bilirubin, a degradation product occurring in the normal catabolic pathway that breaks down heme in vertebrates (PubMed:17187418, PubMed:18004206, PubMed:19830808). Also catalyzes the glucuronidation the isoflavones genistein, daidzein, glycitein, formononetin, biochanin A and prunetin, which are phytoestrogens with anticancer and cardiovascular properties (PubMed:18052087, PubMed:19545173). Involved in the glucuronidation of the AGTR1 angiotensin receptor antagonist losartan, a drug which can inhibit the effect of angiotensin II (PubMed:18674515). Involved in the biotransformation of 7-ethyl-10-hydroxycamptothecin (SN-38), the pharmacologically active metabolite of the anticancer drug irinotecan (PubMed:12181437, PubMed:18004212, PubMed:20610558).Isoform 2Lacks UGT glucuronidation activity but acts as a negative regulator of isoform 1.
Alternative names
UDP-glucuronosyltransferase 1A1, UGT1A1, Bilirubin-specific UDPGT isozyme 1, UDP-glucuronosyltransferase 1-1, UDP-glucuronosyltransferase 1A isoform 1, hUG-BR1, UDPGT 1-1, UGT1*1, UGT1-01, UGT1.1, UGT1, GNT1, UGT1A1
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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.
Alternative names
UDP-glucuronosyltransferase 1A1, UGT1A1, Bilirubin-specific UDPGT isozyme 1, UDP-glucuronosyltransferase 1-1, UDP-glucuronosyltransferase 1A isoform 1, hUG-BR1, UDPGT 1-1, UGT1*1, UGT1-01, UGT1.1, UGT1, GNT1, UGT1A1
Key facts
- Detection method
Fluorescent
- Sample types
Tissue, Suspension cells, Microsomes, Adherent cells
- Assay type
Enzyme activity (quantitative)
- Assay Platform
Microplate
- Sensitivity
= 0.1 mU/well
Storage
- Shipped at conditions
Blue Ice
- Appropriate short-term storage conditions
-20°C
- Appropriate long-term storage conditions
-20°C
- Storage information
-20°C
Notes
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.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
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.
- Associated diseases and disorders
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.
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4 product images
Functional Studies - UGT Activity Assay / Ligand Screening Kit (Fluorometric) (ab273331)
Typical standard curve – data provided for demonstration purposes only. A new standard curve must be generated for each assay performed.
UGT Substrate Standard Curve.
Functional Studies - UGT Activity Assay / Ligand Screening Kit (Fluorometric) (ab273331)
Reaction kinetics of fluorescent substrate glucuronidation in donor-pooled human liver microsomes (HLMs, 0.0625 mg/mL) at 37°C and inhibition of UGT activity in HLMs by the isozyme-selective ligands propofol (UGT1A9-selective) and zidovudine (UGT2B7-selective), as well as the non-selective UGT ligand diclofenac. For the blank reaction condition, vehicle (Assay Buffer) was substituted for the cofactor UDPGA.
Functional Studies - UGT Activity Assay / Ligand Screening Kit (Fluorometric) (ab273331)
Specific Activity in Microsomes.
Specific UGT activity in pooled human and rat liver microsomes (mean ± SEM of 3 replicates).
Functional Studies - UGT Activity Assay / Ligand Screening Kit (Fluorometric) (ab273331)
Dose-response curve for UGT inhibition by diclofenac in HLMs. Percent activity was calculated for each concentration by comparison to activity of reaction containing vehicle only. IC50 values were derived by 4-parameter logistic curve fitting with each point representing the mean ± SEM of at least 3 replicates. Assays were performed according to the kit protocol.
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