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  5. Model TREX1 - Assay System

Model TREX1 -Assay System

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The TREX1 Assay System includes TREX1 enzyme, DNA substrate, assay plates, and buffer components required for a TREX1 reaction to produce dAMP. The Transcreener dAMP Exonuclease Assay (sold separately) measures dAMP to determine enzymatic activity. These measurements allow researchers to screen compound libraries for modulators to help find new disease therapies. The Transcreener dAMP Exonuclease Assay is not included in the assay system but is available for separate purchase through BellBrook Labs. For dAMP assay ordering information, please view Transcreener dAMP Exonuclease Assay.

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Robust Assay Yields Quality Data

The TREX1 Assay System used with the Transcreener dAMP assay produces robust results amenable to HTS. Z’ measurement using optimized TREX1 assay conditions (n=16). Z’ = 0.88.

Screen for Inhibitors & Perform SAR

The TREX1 Assay System used with the Transcreener dAMP assay is designed to screen compound libraries in a high throughput format to identify potential inhibitors. Follow-up SAR can also be performed using the assay to determine inhibitor potency with ease.

Detection of dAMP to Measure TREX1 Activity - FP Readout

The enzyme buffer includes 10 mM Tris pH 7.5, 7.5 mM MgCl2, 0.005% BSA, and 0.01% Brij-35. Here we use 35 nM ISDna. The Detection mix included 20 μg/mL dAMP antibody, 2 nM dAMP Alexa 633 tracer, and 1X Stop & Detect Buffer B in 10 μL. The TREX1 enzyme reaction took place for 60 minutes at 30°C. The Detection Mix was added and incubated for 90 minutes at room temperature and read with a CLARIOstar plate reader. Linearity is shown here when raw data is converted to dAMP formed using a standard curve.

Overview of Transcreener`s dAMP Exonuclease Assay (Sold Separately)
The Transcreener dAMP Exonuclease Assay directly measures dAMP produced during the biological functions like the degradation of DNA. Transcreener dAMP technology uses polyclonal mouse antibodies that selectively bind to dAMP and a far-red fluorescent tracer. dAMP produced displaces the tracer, changing the fluorescent properties and providing a readout.

It is an easy-to-use, mix-&-read, HTS-ready assay that yields robust results. All you need to do is perform the enzyme reaction, add Transcreener reagents, and read your plates. It is available with an FP fluorescent readout.

BellBrook`s Recombinant TREX1 Enzyme

Active, human Three Prime Repair Exonuclease 1 enzyme, His-tagged protein expressed & purified. The enzyme is thoroughly validated with Transcreener dAMP Exonculease Assay Kit. For specific activity, please refer to the Certificate of Analysis for each individual enzyme lot. Purity is >90% in a 50 mM Tris, 500 mM NaCl, 20% glycerol buffer composition (pH 8.0). Currently used with the Transcreener assay at concentrations between 0.1-1.2 nM.