Learn More About the Method Used for the NUDT5 Assay.

Discover Novel Inhibitors with the Transcreener AMP2 NUDT5 Assay

The Transcreener AMP² NUDT5 Assay directly measures the AMP produced by NUDT5 (also known as nudix hydrolase 5 or NUDIX5). By measuring the production of AMP researchers can effectively determine the activity of the NUDT5 enzyme. The assay provides a powerful tool to screen entire compound libraries for NUDT5 modulators to help find new therapeutics for diseases such as cancer.

The kit comes complete with all the detection reagents required to measure activity. NUDT5 enzyme and ADPR are not included in the inhibitor screening assay kit. Please contact us for questions related to acquiring the enzyme.

NUDT5 as a Drug Target

NUDIX hydrolases are a superfamily of enzymes that hydrolyze modified nucleoside diphosphates.  NUDT1 (also known as MTH1) is one of the most well-studied NUDIX hydrolases, having been linked to a variety of disorders including Parkinson's disease and cancer. However, NUDT5 has lately attracted interest due to its link to some human malignancies. NUDT5 hydrolyzes 8-OH-dGDP, 8-OH-dADP, and ADP-ribose (ADPR) as well as other substrates.

High amounts of NUDT1 and NUDT5 have been found in non-small-cell lung cancer (NSCLC) cell lines and tissues, and greater levels of both proteins in NSCLC patients were linked to tumor metastasis.  NUDT5 expression has been shown to be elevated in human breast cancer cell lines and breast tumor tissues.  Patients with high NUDT5 expression profiles also exhibit significantly poorer prognoses than those with lower NUDT5 levels.

Direct Detection of AMP to Measure NUDT5 Enzymatic Activity

NUDT5 Assay FP

Applications

  • Measure Enzymatic Activity of NUDT5
  • Screen Compound Libraries for NUDT5 Inhibitors
  • Quantify Inhibitor Potency
  • Inhibitor Selectivity Profiling

Features

  • Direct detection of unlabeled AMP
  • Easy to use, homogenous, one-step format
  • Robust Assay Z’ > 0.7 under initial velocity conditions
  • Far-red fluorescent readouts minimize compound interference
  • A safe, non-radioactive method
  • Available in FP or TR-FRET readouts

Easy-to-Use, Mix-and-Read, HTS-Ready Assay

The Transcreener AMP² NUDT5 Assay is in a simple, HTS-ready, mix-and-read format. Run your enzyme reaction, add Transcreener reagents, and read your plates. The assay is compatible with 96, 384, and 1536-well formats.

Detection of AMP Under NUDT5 Initial Velocity

The assay demonstrates linearity when raw data is converted to AMP using a standard curve. NUDT5 can use different substrates.  ADPR at 100 μM starting concentration is shown here under initial velocity conditions. The enzyme reaction occurred for 60 minutes at room temperature.

Human NUDT5 Enzyme Titration

Linear Response

Screen for Inhibitors & Perform SAR

Transcreener Assays are designed for screening compound libraries in a high throughput format. Follow up SAR can also be performed using the assay to determine inhibitor potency with ease.

Dose-Response

NUDT5 Assay Dose Response with TH5427

IC50 = 4.66 nM

Robust Assay Yields Quality Data

Z’ measurements using optimized NUDT5 reaction conditions indicate a robust assay.  Quality data like this is vital for sizeable high throughput screens that are difficult to complete due to massive sample quantity. Z' shown here = 0.91

NUDT5 Assay Z Prime Measurement

Far-Red FP & TR-FRET Readouts Validated on Major Multimode Readers

Supplier Instrument FP Assays
TR-FRET Assays
bioteklogo TriStar²S LB 942 in review validated
Mithras² LB 943 in review validated
bioteklogo Cytation™ 5 validated validated
Cytation™ 3 validated validated
Cytation™ 1 validated validated
Synergy™ H1 validated validated
Synergy™ 2/H4/4 validated validated
Synergy™ HTX not capable not capable
Synergy™ Neo 2 validated validated
BMGLABTECH Logo POLARstar® Omega validated validated
FLUOstar® Omega not capable validated
PHERAstar® FSX validated validated
PHERAstar® Plus/FS validated validated
CLARIOstar® /Plus validated validated
MDS AT logo Analyst® GT/HT validated validated
Gemini® XPS/EM not capable not capable
SpectraMax® M2/M2e not capable not capable
SpectraMax® M5/M5e/FlexStation® 3 not capable validated
SpectraMax® Paradigm validated contact us
SpectraMax® iD3/iD5 in review validated
perkinElmerLogo EnVision®/EnVision® Xcite validated validated
tecanLogo Infinite® M1000/M1000Pro/Safire2™ validated validated
Infinite® M200 not capable not capable
Infinite® F500 validated validated
Infinite® F200/Ultraevolution contact us contact us
Spark™ 10M validated validated

What's Included

What You Will Need

NUDT5 Assay Services

Interested in moving your program forward, but don't want to bring an assay in-house? Our scientists can help! BellBrook scientists will use their extensive biochemistry and enzymology expertise to work with you and accelerate your NUDT5 program.

Assay Development Services

Lead Discovery Services Include:

  • Inhibitor Screening – To identify or confirm activity with the target.
  • Inhibitor Potency Profiling – Dose-response with target and/or related proteins. Fast IC50 results.
  • Mechanism of Action Studies – Kinetic analysis to define the mode of inhibition.

Contact Us to Learn More

Please fill out the form below. We will respond quickly to get the conversation moving and learn how we can help. We keep things discrete, confidential, and professional.

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