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 per...
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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 - Benefits
- 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.
Applications
- Measure native MAO activity in various cell lines.
- Measure recombinant MAO activity.
- Screen drugs and chemical entities for their capacity to modulate MAO activity in native or recombinant fractions.
Patents - Disclaimers
Patent Pending.
U.S. Pat. Nos. 6,602,677, 7,241,584 and 8,030,017 and other patents and patents pending.
The method of recombinant expression of Coleoptera luciferase is covered by U.S. Pat. Nos. 5,583,024, 5,674,713 and 5,700,673.
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