Science Catalog 2012 Life Cell Signaling MAO-Glo™ Assay Systems Product MAO-Glo™ Assay MAO-Glo™ Assay with MAO-A Available Separately MAO-A For Research Use Only. Not for Use in Diagnostic Procedures. Description: The MAO-Glo™ Assay provides a homogeneous luminescent method for measuring monoamine oxidase (MAO) activity from recombinant and native sources and for testing the effects of test compounds on MAO activity. The MAO-Glo™ Assay is performed by incubating the MAO enzyme source with a luminogenic MAO substrate. The substrate of the MAO-Glo™ Assay is a derivative of beetle luciferin. Upon reaction with MAO, the derivative is converted into luciferin, which in turn reacts with luciferase to produce light. The amount of light produced is directly proportional to the activity of MAO. After the MAO reaction has been performed, the reconstituted Luciferin Detection Reagent is added. The reagent simultaneously stops the MAO reaction and initiates a stable glow-type luminescent signal with a half-life greater than 5 hours. This eliminates the need for strictly timed luminescent detection. The MAO-Glo™ Assay with MAO-A contains human recombinant MAO-A enzyme expressed in yeast. The kit is very well suited for the rapid assessment of potential inhibition of MAO-A by new chemical entities and can be used for higher throughput applications such as primary screening. The MAO-A enzyme is also available separately. The MAO-Glo™ Assay includes a luminogenic MAO substrate, two MAO Reaction Buffers (one that can be used with either MAO A or MAO B enzyme and one that is designed specifically for MAO B), a lyophilized Luciferin Detection Reagent and the Luciferin Detection Buffer. The user supplies the sample material containing MAO. Protocols are configured for multiwell plate formats but easily can be adapted for single-tube applications. Features: • Complete Solution: The MAO-Glo™ Assay with MAO-A contains monoamine oxidase A enzyme for convenient assessment of the effects of new chemical entities on MAO-A activity. • Speed: The luminescence format eliminates the need for time-consuming analyses such as HPLC. • Simplified Method: The simple “add and read” protocol makes the assay amenable to high-throughput screening in multiwell plates. • Greater Sensitivity: Less MAO enzyme is required in these assays than in typical HPLC or fluorometric methods because of the enhanced sensitivity. • No Fluorescence Interference: Luminescent output eliminates interference from fluorescent test compounds. • Stable Signal: “Glow-type” luminescence provides a stable signal with a half-life of greater than 5 hours. This eliminates the need for strictly timed luminescent detection. Storage Conditions: Store MAO-A enzyme (Cat.# V1452) at –70°C. Store all other components at –20°C protected from light. Protocol MAO-Glo™ Assay Technical Bulletin Part# TB345 Size Cat.# Price (Fr) 200 assays V1401 192.00 1,000 assays V1402 768.00 1,000 assays V1560 1029.00 500 µl V1452 315.00 UGT Activity Assays Product UGT-Glo™ Assay UGT-Glo™ UGT1A1 Screening System UGT-Glo™ UGT2B7 Screening System Size Cat.# Price (Fr) 200 assays V2081 468.00 1,000 assays V2082 1872.00 200 assays V2120 843.00 1,000 assays V2121 3372.00 200 assays V2130 843.00 1,000 assays V2131 3372.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: The UGT-Glo™ Assay is a luminescent method for measuring UDP glucuronosyltransferase (UGT) activity. The UGT-Glo™ Assay measures UGT activity from a variety of sources, such as microsomes containing recombinantly expressed enzymes or microsomal preparations derived from mammalian tissues, and to test the effects of various chemicals on UGT activity. The assay involves incubating UGT with a proluciferin substrate; a portion of the substrate gets conjugated with UDP, while the remainder is unmodified. Upon the addition of D-Cysteine, the unconjugated proluciferin is converted into luciferin and, in a coupled reaction with luciferase/luciferin, is converted into light. Conjugated proluciferin remains intact and does not contribute to the luminescence. Thus, the signal generated is inversely correlated with UGT activity present in the sample. The UGT-Glo™ Assay contains two proluciferin substrates: the UGT Multienzyme Substrate, which is compatible with a wide range of UGTs, and the UGT1A4 Substrate, which reacts specifically with UGT1A4. The kit also contains Luciferin Detection Reagent and Reconstitution Buffer, UGT Buffer, D-Cysteine and UDPGA. The UGT-Glo™ Screening Systems contain the above reagents as well as the respective UGT isoforms and control membranes. Features: • Speed: The luminescent format eliminates the need for time-consuming analyses such as HPLC and LC/MS. • Simplified Method: The simple “add and read” protocol makes the assay amenable to higher throughput screening in multiwell plates. • Sensitive: Allows researchers to use less enzyme and scale down reaction volumes, which saves on reagent costs. Storage Conditions: Store UGT enzymes and Control Membranes at –70°C. Store remaining components at –20°C. Protocol UGT-Glo™ Assay Technical Bulletin Part# TB551 24 For complete and up-to-date product information visit: www.promega.com/catalog Pagina 27
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